new biorapid product
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new biorapid product
14th June 2010 286/MKT/BR/20 NEW biorapid MONONUCLEOSIS 25 Test NEW BIORAPID PRODUCT We are happy to announce the launch of a new product of the biorapid family based on immunochromatography technology: - biorapid MONONUCLEOSIS 25 Test code 3000-1027 This product is intended for the qualitative detection of heterophile antibodies to Infectious Mononucleosis. biorapid MONONUCLEOSIS completes the very successful biokit range of products for IM: Monogen and Color-Mono. Find enclosed complete product information that we hope helps you in the launch of the assay. Good sales and good luck! Index: biorapid MONONUCLEOSIS - General information on EBV Diagnosis biorapid MONONUCLEOSIS Assay procedure Performance Competitors Features and Benefits Conclusion Package insert Page 2 of 13 Infectious Mononucleosis General information on the Epstein-Barr virus: The Epstein-Barr virus (EBV) was first discovered in 1964 as the cause of infectious mononucleosis. (IM). This disorder is usually an acute, benign and self-limiting lymphoproliferative condition that is usually subclinical in children and a mild condition in adults. The EBV is also the cause of nasopharyngeal carcinoma and neoplasms of the thymus, parotid gland and larynx. Burkitt’s lymphoma, a malign tumour of the lymphoid tissue occurring mainly in African children, is also caused by EBV. Epstein-Barr virus is a gamma-herpesvirus (see figure 1). The EBV has a very limited host range and tissue tropism: Human B-lymphocytes and epithelial cells of the oropharynx and nasopharynx. EBV is widely disseminated. It is estimated that 95% of the world population is exposed to the virus, which makes it the most widespread virus known to man. EBV appears to be transmitted primarily by close contact with infectious oral-pharyngeal secretions, however the virus has been reported to be transmitted by blood transfusion and transplacental routes. However, under ordinary conditions, transmission of the virus through transfusion or transplacental exposure is unlikely. In addition, EBV-associated post-transplantation lymphoproliferative disease develops in 1% to 10% of organ transplant recipients. The frequency of seronegative patients is nearly 100% in early infancy, but declines more or less rapidly with increased age and depending on socio-economic conditions, to less than 10% in young adults. After primary exposure, a person is considered to be immune and generally no longer susceptible to new re-infections. In Western society, primary exposure to EBV occurs in two waves. Approximately half of the population is exposed to the virus before the age of 5 years. A second wave of seroconversion occurs during late adolescence, between 15 to 24 years of age. More than 90% of EBV-infected individuals intermittently shed the virus for life even when totally asymptomatic. Children can acquire the virus at an early stage by sharing contaminated drinking glasses and generally undergo subclinical disease. Saliva sharing between adolescents and young adults often occurs by kissing, thus the nickname of “kissing disease”. In these individuals the disease may go unnoticed or be present in varying degrees of severity as infectious mononucleosis (IM). Clinical symptoms The cardinal clinical features of the mononucleosis syndrome are fever, sore throat, malaise, and fatigue, accompanied by signs of tonsillopharyngitis and lymphadenopathy. There is frequently a prodromal period of 2 - 5 days consisting of malaise and fatigue, prior to the onset of the full syndrome. The adenopathy in mononucleosis most prominently involves the anterior and posterior cervical lymph nodes, but diffuse adenopathy also often occurs. Splenomegaly develops in the first 3 weeks of illness in about 50% of cases. The adenopathy and organomegaly are often most prominent in the second to fourth weeks of illness. Other frequently observed clinical signs include: rash, usually erythematous and maculopapular, soft palate petechiae, bilateral edema of the eyelids, and jaundice. It is well established that ampicillin will induce a typical rash in virtually all patients with mononucleosis. Page 3 of 13 Infection with EBV has been shown to be very common in small children. By the age of 4, as many as 70% of children will have acquired the antibody. The clinical manifestations in children under 4 years of age typically entail more Patients with infectious mononucleosis typically demonstrate an absolute lymphocytosis (greater than 50%), prominent atypical lymphocytosis (often greater than 10- 20%), a positive test for Paul- Bunnell heterophile antibodies, and mild to moderate elevation of liver enzymes. A diagnosis made on the basis of the clinical record and symptomatology alone is difficult. Numerous cases have been cited in which IM has been misidentified with other non-related viral and bacterial diseases. For this reason, blood and serum tests are very helpful in diagnosis. Diagnosis of Epstein-Barr virus infection Diagnosis of EBV could be divided into non-specific and specific methods. Heterophile antibodies The immunitary system of infected patients produces heterophile antibodies, most of them IgM. These antibodies do not react directly to EBV antigens but even appear a little bit earlier than EBV specific IgM antibodies. The heterophile antibody that appears in the EBV infection also has the capability of reacting to a glycoprotein present on the surface of sheep erythrocyte antigen. This antigen was discovered by Paul-Bunnell in 1932. Heterophile antibody assays are very useful as screening tests as they detect the very early primary infection. biokit has a well known rapid test based on the Paul-Bunnell antigen, the Monogen (Code number 3000-1001) and a rapid test based on slide haemagglutination, Color-mono (Code number 3000-1005). Elisa tests Enzyme immunoassay tests are based on purified proteins, recombinant antigens, or synthetic peptides. There are specific tests for detection of all EBV antigens for IgG or IgM such as VCA, EBNA or Early Antigens. The Elisa test provides the best combination of sensitivity, specificity and user-friendliness for confirmation and followup. Immunofluorescence assays These are considered as a reference method for EBV diagnosis.. The assays are also based on recombinant antigens or synthetic peptides. They use microscope slides as a support for the solid phase and anti IgG or IgM conjugate labelled with a fluorescent marker. These methods require highly skilled staff to read and interpret the results. Immunoblot Enzyme immunoassay tests based on recombinant antigens that use a nitro-cellulose strip as a support for the solid phase. The strip incorporates all the antigens separately, thus allowing the assay to determine the presence of specific antibodies to these antigens in the patient samples. Page 4 of 13 biorapid MONONUCLEOSIS biorapid MONONUCLEOSIS is a rapid, one-step qualitative screening test for the detection of Infectious Mononucleosis heterophile antibodies in serum, plasma or whole blood. The test is able to specifically detect heterophile antibodies in the early phase of Infectious mononucleosis. biorapid MONONUCLEOSIS is a rapid qualitative one-step immunochromatographic assay. The test device includes a sample well (S), a conjugate containing colloidal gold particles coated with highly purified Paul-Bunnell antigen, and a chromatographic membrane strip where Paul-Bunnell antigen has been immobilized to act as a capture means. Sensitivity has been adjusted to mach that provided by Monogen (latex based assay), a worldwide reference for IM diagnosis. Method The sample is introduced into the sample well (S) and allowed to migrate along the attached membrane strip. If IM heterophile antibodies are present in the sample, they will bind to the conjugate forming an immunocomplex. The complex continues to migrate further along the membrane where it will be captured by the Paul-Bunnell antigen immobilized in the Test zone (T), leading to the formation of a coloured line. The remaining conjugate will bind to the control zone (C) forming a second coloured line indicating correct performance of the test. If IM heterophile antibodies are present in the sample, two coloured lines will appear in the reading window and indicate a positive result. If IM heterophile antibodies are absent, only the Control line will appear and a negative result is indicated. If no lines or only the Test line appears, the test is invalid. Material provided - 25 biorapid MONONUCLEOSIS test devices. 1 x 2 ml diluent buffer. 1 x 1 ml positive control. 1 x 1 ml negative control. 25 disposable pipettes. Page 5 of 13 Sample collection biorapid MONONUCLEOSIS can be used for serum, plasma (EDTA, heparin and citrate) or Whole Blood (EDTA, heparin and citrate). Specimens that contain particulate material may give inconsistent results and should be clarified before testing by high speed centrifugation (10,000 rpm) for 10 minutes. Storage and stability biorapid MONONUCLEOSIS should be stored at 2-8°C. The stability of biorapid MONONUCLEOSIS is that stated on the kit label if stored under the above conditions. Test devices and diluent buffer can be stored separately at any temperature between 225°C. Quality control The blue line at the location of the control zone is for manufacturing control purposes only and does not interfere with performance of the test. Any blue dye that may remain in the window at reading time does not interfere with the results. The Control line (C) is an internal procedural control. This line will always appear if the test has been performed correctly and the test device is functioning properly. Use the controls included in the kit to check the performance of the biorapid MONONUCLEOSIS assay. If the expected results are not obtained, do not use the kit. Page 6 of 13 Assay procedure FOR SERUM, PLASMA AND CONTROLS: - Allow test devices and samples to reach room temperature before use.. Remove the test device from the pouch and place it on a flat surface. Label the test device with the patient’s name or identification number. Hold the pipette in a vertical position and dispense 3 drops (75 µl) of serum or plasma into the sample well (S). For the positive and negative controls, hold the dropper in a vertical position and dispense 2 drops into the sample well (S). Immediately add 2 drops of the diluent buffer holding the dropper in a vertical position. Read the result after 5 minutes and then discard the test device. Do not read a test result after more than 5 minutes have elapsed. FOR WHOLE BLOOD: - Allow test devices and samples to reach room temperature before use.. Remove the test device from the pouch and place it on a flat surface. Label the test device with the patient’s name or identification number. Hold the pipette in a vertical position and dispense 2 drops (50 µl) of blood into the sample well (S). Immediately add 3 drops of the diluent buffer holding the dropper in a vertical position. Read the result after 5 minutes and then discard the test device. Do not read a test result after more than 5 minutes have elapsed. DO NOT INTERPRET THE TEST AFTER 15 MINUTES Interpretation of the results POSITIVE If two coloured lines (Test and Control) are visible, the test result is positive. Any visible colour on the Test line (T) should be interpreted as a positive result even if it has less intensity than the Control line (C). NEGATIVE If only the Control line (C) is visible, the test result is negative. INVALID If no distinct lines appear or if only the Test line is visible, the test is invalid. The sample should be retested using another test device. Page 7 of 13 POSITIVE biokit NEGATIVE biokit INVALID biokit C C C T T T Limitations of the procedure - - The results should be interpreted in light of the clinical, haematological, and serological information of the patient. Occasionally detectable levels of heterophile antibodies are late in developing in symptomatic IM patients.. If symptoms persist, the assay should be repeated within a few days. Some patients, especially children and adolescents, may remain persistently negative. It has been reported that only 80 to 90% of adults and less than 50% of young children develop heterophile antibodies. In some individuals, detectable levels of heterophile antibodies may persist for months and, more rarely, for years. Expected values Different studies of the presence of IM heterophile antibodies in blood donors show that the incidence of the disease ranges from 0.9 to 1.7% of the population. As the presence of heterophile antibodies indicates a relatively recent infection, these results suggest that the true incidence of the disease is higher than the number of diagnosed cases. Page 8 of 13 Performance characteristic A total of 622 samples (296 serum samples, 261 plasma samples, and 65 whole blood samples) were evaluated both in an internal and external study using biorapid MONONUCLEOSIS and another commercially available immunochromatographic (ICT) test: Genzyme OSOM Mono Test. The Genzyme kit is a FDA approved market leader in the United States. All samples were confirmed as positive or negative by a commercially available latex particle agglutination test for the qualitative detection of IM heterophile antibodies. Results Serum samples biorapid MONONUCLEOSIS Genzyme Mono Test + - + 75 0 - 1* 220 (*) The discrepant sample was negative for a latex agglutination test. Plasma samples biorapid MONONUCLEOSIS Genzyme Mono Test + - + 52 0 - 1* 208 (*) The discrepant sample was negative for a latex agglutination test. Whole blood samples biorapid MONONUCLEOSIS Genzyme Mono Test + - + 15 0 - 0 50 All samples biorapid MONONUCLEOSIS Genzyme Mono Test + - + 142 0 - 2 478 When compared to Genzyme Mono Test, biorapid MONONUCLEOSIS showed a sensitivity of 100% (142/142) and a specificity of 99.6% (478/480). The overall agreement between the two ICT tests was 99.7%. Comparison of biorapid MONONUCLEOSIS to Biokit Monogen biorapid MONONUCLEOSIS Page 9 of 13 MONOLATEX + - + 120 0 - 9** 428 When compared to the Biokit MONOLATEX agglutination test, the biorapid MONONUCLEOSIS test showed a sensitivity of 100% (120/120) and a specificity of 97.9% (428/437). The overall agreement was 98.4%. Reproducibility A reproducibility evaluation of the biorapid MONONUCLEOSIS test was conducted at 3 external laboratories where testing was performed by personnel with diverse educational backgrounds. At each site, testing was performed on randomly coded samples, 3 positive and 3 negative, for 3 days and in triplicate. The results obtained showed a 100% agreement with the expected results. Page 10 of 13 Competitors Competitors on lateral flow Two main competitors may be found in lateral flow technology: - OSOM Mono Test, Genzyme - Clearview IM, Inverness The following table summarises all the characteristics and stated performance: Test Name No. of Tests Device shape Negative Control Positive Control Sample Diluent Capillary tubes & bulb for blood collection Disposable pipettes Incubation time Storage Declared Sensitivity Declared Specificity Comments BIOKIT Biorapid Mononucleosis 25 T Plastic cassette GENZYME OSOM Mono Test 25 T Only Strips INVERNESS Clearview IM 20 T Plastic cassette YES YES YES NO YES YES YES YES NO NO YES NO YES NO 5 min. 5 min. YES 5 min. Serum/Plasma 15 min. Whole Blood RT: Device + Diluent 2-8 °C, Kit controls RT RT 100% 100% 95.5% (blood) 98.5% (Serum/Plasma) 95.9% 100% 99.5% vs. lateral flow 97.9% vs. Latex + Controls included + Sharp clear bands + - - Controls included Poor readability, faint bands Strip assay: Strip must be dipped in the sample tube. More manipulation Potential risk of sample contamination + - Clear bands Does not include controls Page 11 of 13 Internal evaluation vs. competitors A total of 21 clinical samples, characterized as positive by biokit Monogen but with a different degree of reactivity, were evaluated using biorapid MONONUCLEOSIS, Genzyme OSOM Mono Test and Clearview IM. Results biorapid MONONUCLEOSIS detected 18 out of 21 samples. Genzyme OSOM Mono Test detected 15 out of 21 samples. Clearview IM detected 16 out of 21 samples. biorapid MONONUCLEOSIS Monogen + + - 18 3 Genzyme OSOM Mono Test Monogen + + - 15 6 Clearview IM Monogen + + - 16 5 The biorapid MONONUCLEOSIS showed the best correlation with the biokit Monogen, considered as a worldwide reference for the detection of heterophile antibodies against Infectious Mononucleosis. Page 12 of 13 Features and benefits biorapid MONONUCLEOSIS Features Benefits Biokit’s own Paul Bunnell antigen - The best antigen on the market as demonstrated by Monogen and Color-Mono. Correlation with Monogen Official approvals - Excellent sensitivity Excellent specificity CE mark US FDA Positive and Negative Controls included - Full confidence in the results obtained Clearview does not include controls - Doctor’s Office Test Easiest sample to be taken No Sample preparation No lab tubes to collect serum or plasma Cost savings Also suitable for the typical samples used in any laboratory Objective results Sharp bands No background No need for staff trained people in reading agglutination tests Blood Finger tip sample Serum or plasma sample - Easy reading Room temperature storage for test devices and diluent - No extra refrigeration equipment required More room in refrigerators Long shelf-life - Simplifying distributor and end-users maintaining a correct inventory Easy manipulation compared to strip tube assays Strip tube assay needs sample aliquot to dip the strip in. Potential risk of sample contamination Plastic device - Conclusion biorapid MONONUCLEOSIS is manufactured by one of the most expert companies in IM diagnosis. Monolatex and Color-Mono are worldwide references in quality according to different international publications. As demonstrated in the evaluations, biorapid MONONUCLEOSIS matches the performance of the Biokit reference assay, Monolatex, and the US market leader for lateral flow IM detection, the Genzyme Mono Test. Apart from performance, the biorapid MONONUCLEOSIS test shows important advantages vs. competitors: • Vs. Clearview: biorapid includes positive and negative controls whereas Clearview does not. • Vs. Genzyme: The biorapid assay is a practical plastic device. The Genzyme is a strip-tube assay. The strip must be dipped in the sample tube. Page 13 of 13 biorapid MONONUCLEOSIS READ HIGHLIGHTED CHANGES REF 3000-1027 3800-3038 05/10 Amostras de sangue total Teste comparativo Total amostras biorapid MONONUCLEOSIS biorapid MONONUCLEOSIS + - + 15 0 - 0 50 One-step immunoassay for the detection of infectious mononucleosis heterophile antibodies in serum, plasma or whole blood. biorapid MONONUCLEOSIS + Teste comparativo Summary - + 142 0 - 2 478 Ao ser comparado com um teste imunocromatográfico (ICT) comercial, o teste biorapid MONONUCLEOSIS mostrou uma sensibilidade de 100% (142/142) e uma especificidade de 99,6% (478/480). A concordância total entre os dois testes foi de 99,7%. Ao ser comparado com um teste comercial de aglutinação de partículas de látex, o teste biorapid MONONUCLEOSIS mostrou uma sensibilidade de 100% (120/120) e uma especificidade de 97,9% (428/437). A concordância total entre os dois testes foi de 98,4%. Infectious mononucleosis (IM) is an acute infectious disease caused by the Epstein-Barr virus (EBV). In 1932, Paul and Bunnell reported that the serum of patients with IM develop high titers of heterophile antibodies to sheep erythrocytes. Also were described agglutinins to red blood cells from other mammals. The proteins responsible for this agglutination are glycoproteins from red cell membranes called Paul-Bunnell antigen by several authors. Studies made on these glycoproteins show that those purified from bovine red blood cells are the most sensitive to IM heterophile antibodies. Heterophile antibodies to sheep erythrocytes may also be detected in sera from normal people, individuals who have received injections of serum, and others. Traditionally the IM heterophile antibodies have been distinguished from other heterophile antibodies by a “differential” absorption test with bovine red blood cells and guinea pig kidney tissue. The use of highly purified Paul-Bunnell antigen provides a simple method with improved sensitivity for the specific detection of heterophile antibodies associated with IM. Interferências Principle Para estudar possíveis interferências foram analizadas amostras com risco potencial de produzir reações falsopositivas, como 10 amostras de soro positivas de anticorpos IgM anti-toxoplasma, 10 positivas de anticorpos IgM anti-CMV e 10 positivas para fator reumatóide. Todas amostras apresentaram resultados negativos. biorapid MONONUCLEOSIS is a rapid qualitative one step immunochromatographic assay. The test device includes a sample well (S), a conjugate containing colloidal gold particles coated with highly purified Paul-Bunnell antigen, and a chromatographic membrane strip where Paul-Bunnell antigen has been immobilized to act as a capture. Reprodutibilidade The sample is added into the sample well (S) and allowed to migrate along the attached membrane strip. If IM heterophile antibodies are present in the sample, they will bind to the conjugate forming an immunocomplex. The complex continues to migrate further along the membrane where it will be captured by the Paul-Bunnell antigen immobilized on the test zone (T), leading to the formation of a coloured line. Remaining conjugate will bind to the control zone (C) forming a second coloured line indicating the proper performance of the test. Uma avaliação da reprodutibilidade do teste biorapid MONONUCLEOSIS foi realizada em 3 laboratórios externos onde a prova foi realizada por pessoal com diferente nível de treinamento. Em cada laboratório, o teste se realizou com amostras rotuladas aleatoriamente, 3 positivas e 3 negativas, durante 3 dias e por triplicado. Os resultados obtidos apresentaram 100% de concordância com os resultados esperados. Bibliografia Ver “References” do texto inglês. If IM heterophile antibodies are present in the sample, two coloured lines will appear in the reading window and indicate a positive result. If IM heterophile antibodies are absent, only the Control line will appear and a negative result is indicated. If no lines or only the Test line appears, the test is invalid. Components REF 3000-1027. - TEST 25 biorapid MONONUCLEOSIS test devices. - DIL 1 x 2 ml diluent buffer.Tris buffer with additives. - CONTROL + 1 x 1 ml positive control. Rabbit IgG anti-Paul-Bunnell antigen diluted in a buffer. Contains < 0.1% sodium azide. - CONTROL - 1 x 1 ml negative control. Non reactive diluted human serum. Contains < 0.1% sodium azide. - PIPETTES 25 disposable pipettes. Precautions biorapid MONONUCLEOSIS is intended for IN VITRO diagnostic use. Sodium azide may react with lead or copper pipes and plumbing creating highly explosive metal azides. Flush drains with water thoroughly after disposing of the remains of reagents. All human source material used in the preparation of this product was found to be negative for the presence of HIV1/HIV-2 and HCV antibodies, as well as for the hepatitis B surface antigen, using a method approved by the Food and Drug Administration (USA). WARNING: POTENTIALLY BIOHAZARDOUS MATERIAL. Because no test method can offer complete assurance of the absence of infectious agents, this product should be handled with caution. Do not use test devices if the aluminum pouch has been damaged during storage. A new pipette must be used for each sample to avoid contamination errors. Dispose all used materials in a suitable biohazardous waste container. 24 1 Storage The reagents will remain stable through the expiration date shown on the label if stored between 2-8°C. Do not freeze. Test devices and diluent buffer could be stored separately at 2 to 25°C. biokit biokit biokit Sample collection SERUM or PLASMA: Use fresh serum or plasma (EDTA, heparin, citrate). Other anticoagulants should be evaluated before use. Samples can be stored at 2-8°C for 2 days. For longer periods samples should be frozen (-20°C). Avoid repeated freezing and thawing. Homogenize the samples before analysis. Grossly hemolyzed, turbid, or lipemic samples should not be used. Samples that contain particulated material may give inconsistent results and should be clarified by centrifugation before testing. C C C T T T S S S WHOLE BLOOD: Collect whole blood samples using a tube containing anticoagulant (EDTA, heparin, citrate). Other anticoagulants should be evaluated before use. Whole blood samples can be stored at 2-8°C for 2 days. POSITIVO Quality control NEGATIVO The blue line at the location of the control zone (C) is for manufacturing control purposes only and does not interfere with the performance of the test. Any blue dye that may remain in the window at the reading time, does not interfere with the results. Limitações do procedimento The Control line (C) is an internal procedural control. If the test has been performed correctly and the test device is functioning properly this line will always appear. - To check the performance of biorapid MONONUCLEOSIS, use the controls included in the kit. If expected results are not obtained, do not use the kit. Procedure - - SEM VALIDADE Os resultados devem ser interpretados tendo em consideração a informação clínica, hematológica e serológica do paciente. Ocasionalmente níveis detectáveis de anticorpos heterófilos se desenvolvem tardiamente em pacientes sintomáticos de MI. Se os sintomas persistirem se recomenda repetir o teste após uns dias. Alguns pacientes, especialmente crianças e adolescentes, podem persistir permanentemente como negativos. Se publicou que apenas de 80% a 90% dos adultos e menos de 50% dos jovens desenvolvem anticorpos heterófilos. Em alguns indivíduos podem persistir níveis detectáveis de anticorpos heterófilos durante meses e, mais raramente, durante anos. FOR SERUM, PLASMA, and CONTROLS: Valores previstos - Diferentes estudos realizados em doadores de sangue indicam que a presença de anticorpos heterófilos de MI oscila entre 0,9 e 1,7% da população. Dado que a presença destes anticorpos indica uma infecção relativamente recente, os resultados obtidos sugerem que a incidência real da doença pode ser maior que o número de casos diagnosticados. - Allow test devices and samples to reach room temperature before using. Remove test device from the pouch and place it on a flat surface. Label test device with patient name or identification number. Hold the pipette in vertical position and dispense 3 drops (75 µl) of serum or plasma into the sample well (S). For the positive and negative controls, hold the dropper in vertical position and dispense 2 drops into the sample well (S). Immediately after, add 2 drops of the diluent buffer holding the dropper in vertical position. Read the result at 5 minutes, then discard the test device. Do not read a test result after more than 5 minutes. FOR WHOLE BLOOD: - Allow test devices and samples to reach room temperature before using. Remove the test device from the pouch and place it on a flat surface. Label test device with patient name or identification number. Hold the pipette in vertical position and dispense 2 drops (50 µl) of blood into the sample well (S). Immediately after, add 3 drops of the diluent buffer holding the dropper in vertical position. Read the result at 5 minutes, then discard the test device. Do not read a test result after more than 5 minutes. Características funcionais Um total de 622 amostras (296 soros, 261 plasmas e 65 sangue total) foram avaliadas internamente e em uma avaliação externa utilizando o teste biorapid MONONUCLEOSIS e outro teste imunocromatográfico (ICT) comercial. Todas as amostras foram confirmadas como positivas ou negativas mediante um teste comercial de aglutinação de partículas de látex. Resultados Amostras de soro biorapid MONONUCLEOSIS Teste comparativo Interpretation of the results + - + 75 0 - 1* 220 POSITIVE If two coloured lines (Test and Control) are visible, the test result is positive. Any visible colour on the Test line (T) should be interpreted as a positive result even if it has less intensity than the Control line (C). NEGATIVE (*) A amostra discrepante deu resultado negativo com um teste de aglutinação de látex. Amostras de plasma biorapid MONONUCLEOSIS If only the Control line (C) is visible, the test result is negative. + INVALID If no distinct lines appear or if only the Test line is visible, the test is invalid. The sample should be retested using another test device. Teste comparativo - + 52 0 - 1* 208 (*) A amostra discrepante deu resultado negativo com um teste de aglutinação de látex. 2 23 Conservação Os reativos permanecem estáveis até a data de validade indicada na etiqueta, se forem conservados entre 2-8°C. Não congelar. Os dispositivos e o tampão diluente podem ser conservados em separado entre 2 e 25°C. biokit biokit biokit Coleta da amostra SORO OU PLASMA: Usar soro fresco ou plasma (EDTA, heparina, citrato). Outros anticoagulantes devem ser avaliados antes de serem utilizados. As amostras podem ser conservadas por 2 dias entre 2-8°C. Para guardar por um período de tempo mais longo as amostras devem ser congeladas (-20°C). Evitar congelar e descongelar as amostras repetidamente. Homogeneizar as amostras antes de serem analisadas. Não se devem utilizar amostras hemolizadas, turvas ou lipêmicas. As amostras que contêm material particulado podem dar resultados inconsistentes e necessitam ser clarificadas por centrifugação antes de realizar o teste. C C C T T T S S S SANGUE TOTAL: Coletar o sangue em tubos com anticoagulante (EDTA, heparina, citrato). Outros anticoagulantes devem ser comprovados antes de serem utilizados. As amostras de sangue total podem ser conservadas durante 2 dias entre 2 e 8°C. POSITIVE Controle de qualidade Limitations of the procedure A linha azul que se observa na zona Controle (C) é um controle de fabricação e não interfere no funcionamento do teste. Qualquer resto de corante azul que se pudera visualizar na janela no momento de realizar a leitura, não interfere nos resultados. - NEGATIVE INVALID The results should be interpreted in light of the clinical, hematological, and serological information of the patient. Occasionally detectable levels of heterophile antibodies are late in developing in patients symptomatic for IM. If symptoms persist it is recommended to repeat the assay in several days. Some patients, especially children and adolescents, may remain persistently negative. It has been reported that only 80 to 90% of adults and less than 50% of young children develop heterophile antibodies. In some individuals, detectable levels of heterophile antibodies may persist for months and, more rarely, for years. A linha Controle (C) é um controle interno do procedimento. Se o teste foi realizado corretamente e o dispositivo funciona devidamente, esta linha sempre deve aparecer. - Para comprovar o correto funcionamento do biorapid MONONUCLEOSIS utilizar os controles incluídos no kit. Se não se obtêm os resultados esperados, não utilize o kit. Expected values Procedimento Different studies of presence of IM heterophile antibodies in blood donors show that the incidence of the disease ranges from 0.9 to 1.7% of population. As presence of heterophile antibodies indicates a relatively recent infection, these results suggest that the true incidence of the disease is higher than the number of diagnosed cases. PARA SORO, PLASMA E CONTROLES: - Deixar que os dispositivos e as amostras atinjam a temperatura ambiente antes de utilizá-los. Retirar o dispositivo da embalagem e colocá-lo sobre uma superfície plana. Rotular o dispositivo com o nome do paciente ou número de identificação. Manter a pipeta em posição vertical e dosificar 3 gotas (75 µl) de soro ou plasma no pozinho de amostra (S). Para os controles positivo e negativo, manter o conta-gotas em posição vertical e dosificar 2 gotas no pozinho de amostra (S). Imediatamente depois, acrescentar 2 gotas de tampão diluente, mantendo o conta-gotas em posição vertical. Ler o resultado após 5 minutos e descartar o dispositivo. Não fazer a leitura dos resultados em caso de se exceder o período de tempo indicado. Performance characteristics A total of 622 samples (296 serum samples, 261 plasma samples, and 65 whole blood samples) were evaluated internally and in an external study with biorapid MONONUCLEOSIS and another commercially available immunochromatographic (ICT) test. All samples were confirmed as positive or negative by a commercially available latex particle agglutination test for the qualitative detection of IM heterophile antibodies. Results Serum samples PARA SANGUE TOTAL: - Deixar que os dispositivos e as amostras atinjam a temperatura ambiente antes de utilizá-los. Retirar o dispositivo da embalagem e colocá-lo sobre uma superfície plana. Rotular o dispositivo com o nome do paciente ou número de identificação. Manter a pipeta em posição vertical e dosificar 2 gotas (50 µl) de sangue no pozinho de amostra (S). Imediatamente depois, acrescentar 3 gotas de tampão diluente, mantendo o conta-gotas em posição vertical. Ler o resultado após 5 minutos e descartar o dispositivo. Não fazer a leitura dos resultados em caso de se exceder o período de tempo indicado Interpretação dos resultados Comparative test NEGATIVO Em caso de que apenas a linha de Controle (C) adquira cor, o resultado será considerado negativo. 22 - 75 0 - 1* 220 biorapid MONONUCLEOSIS Comparative test SEM VALIDADE Se nenhuma das linhas adquire cor ou apenas a linha de Teste fica colorida, o resultado será considerado sem validade. Portanto, o teste deverá ser repetido utilizando-se outro dispositivo. + + (*) The discrepant sample was negative by a latex agglutination test. Plasma samples POSITIVO Se as duas linhas (Teste e Controle) forem visíveis, o resultado será positivo. Qualquer cor que a linha de Teste (T) adquira, será indicação de resultado positivo, inclusive se a cor for menos intensa que a da linha de Controle (C). biorapid MONONUCLEOSIS + - + 52 0 - 1* 208 (*) The discrepant sample was negative by a latex agglutination test. 3 Whole blood samples biorapid MONONUCLEOSIS biorapid MONONUCLEOSIS Comparative test + - + 15 0 - 0 50 Teste de um só passo para a detecção de anticorpos heterófilos da mononucleose infecciosa em soro, plasma ou sangue total. Sumário All samples biorapid MONONUCLEOSIS Comparative test + - + 142 0 - 2 478 When compared to a commercially available immunochromatographic (ICT) test, biorapid MONONUCLEOSIS showed a sensitivity of 100% (142/142) and a specificity of 99.6% (478/480).The overall agreement between the two ICT tests was 99.7% When compared to a commercially available latex particle agglutination assay, the biorapid MONONUCLEOSIS test showed a sensitivity of 100% (120/120) and a specificity of 97.9% (428/437). The overall agreement was 98.4%. Interferences To study possible interferences, samples with potential risk of producing false positive results were tested, including 10 serum samples positive for toxoplasma IgM antibody, 10 positive for CMV IgM antibody and 10 positive for rheumatoid factor. All samples gave a negative result. Reproducibility A reproducibility evaluation of the biorapid MONONUCLEOSIS test was conducted at 3 external laboratories where testing was performed by personnel with diverse educational backgrounds. At each site, testing was performed on randomly coded samples, 3 positive and 3 negative, for 3 days and by triplicate. Results obtained showed a 100% agreement with the expected results. References 1. Biosafety in Microbiological and Biomedical Laboratories. CDC/NIH manual, 5th Edition, 2007. 2. Davidsohn I. Serologic diagnosis of infectious mononucleosis. JAMA 108: 289-295, 1937. 3. Gray JJ, Caldwell J, Sillis M. The rapid serological diagnosis of infectious mononucleosis. Journal of Infection 25: 39-46, 1992. 4. Linderholm M, Boman J, Juto P, Linde A. Comparative evaluation of nine kits for rapid diagnosis of infectious mononucleosis and Epstein-Barr virus-specific serology. J Clin Microbiol 32: 259-261, 1994. A mononucleose infecciosa (MI) é uma doença infecciosa aguda causada pelo vírus de Epstein-Barr (EBV). Em 1932, Paul e Bunnell observaram que os soros de pacientes com MI continham anticorpos heterófilos contra hemácias de carneiro. Mais tarde também foram descritas aglutininas contra eritrócitos de outros mamíferos. As proteínas responsáveis por esta aglutinação são glicoproteínas de membrana dos glóbulos vermelhos, denominadas antígeno de Paul Bunnell por diversos autores. Os estudos realizados demonstram que o antigênio de Paul-Bunnell purificado de eritrócitos bovinos é mais sensível aos anticorpos heterófilos da MI que o procedente de outros mamíferos. Também se pôde detectar anticorpos heterófilos contra eritrócitos de carneiro diferentes dos da MI em soros de pessoas saudáveis, de indivíduos que receberam injeções de soros, e outros. Tradicionalmente os anticorpos heterófilos da MI têm sido distinguidos de outros tipos através da uma análise de absorção “diferencial” realizada con hemácias bovinas e tecido renal de cobaia. O uso do antígeno de Paul-Bunnell altamente purificado neste teste proporciona um método mais simples e mais sensível para a detecção dos anticorpos heterófilos de MI. Princípio biorapid MONONUCLEOSIS é um teste imunocromatográfico de um só passo. O dispositivo inclui um pozinho para a adição da amostra (S), um conjugado de partículas de ouro coloidal sensibilizadas com antígeno de Paul-Bunnell altamente purificado, e uma membrana cromatográfica na que se imobilizou o antígeno de Paul-Bunnell, que atuará como captor. A amostra se dosifica no pozinho correspondente (S) e começa a migrar ao longo da membrana adjunta. Se a amostra contém anticorpos heterófilos de MI, estes se unirão ao conjugado formando um imunocomplexo. O complexo continua sua migração ao longo da membrana onde será capturado pelo antígeno de Paul-Bunnell imobilizado na zona Teste (T), dando lugar à formação de uma linha colorida. O conjugado restante se unirá à zona Controle (C) formando uma segunda linha colorida, a qual indica o bom funcionamento do teste. Se a amostra contém anticorpos heterófilos de MI, na janela de leitura aparecem duas linhas coloridas indicando um resultado positivo. Se a amostra não contém anticorpos heterófilos de MI, só aparecerá a linha Controle indicando um resultado negativo. Se não aparece nenhuma linha ou se só aparece a linha Teste, o resultado não se considera válido. Componentes 5. Medical Devices Agency Evaluation Report: An evaluation of fourteen commercial kits used to screen for the presence of infectious mononucleosis (MDA/98/63). Medical Devices Agency Evaluation Centre. UK National Health Service. 6. Pozzetto B, Mbida AD, Bourlet T, Grattard F, Bonnevial L. Comparative evaluation of eight commercial tests for the diagnosis of infectious mononucleosis by Epstein-Barr virus-specific or –non specific serology. Serodiagn Immunother Infect Dis 8: 221-223, 1997. 7. Virtanen S. Incidence of infectious mononucleosis antibodies in blood donors. Acta Pathol Microbiol Immunol Scand 56: 53-56, 1962. 4 REF 3000-1027. - TEST 25 dispositivos biorapid MONONUCLEOSIS. - DIL 1 x 2 ml tampão diluente. Tampão Tris com aditivos. - CONTROL + 1 x 1 ml controle positivo. IgG de coelho contra o antígeno de Paul-Bunnell diluído em um tampão. Contém azida sódica < 0,1%. - CONTROL - 1 x 1 ml controle negativo. Soro humano negativo diluído. Contém azida sódica < 0,1%. - PIPETTES 25 pipetas descartáveis. Precauções biorapid MONONUCLEOSIS é para o diagnóstico IN VITRO. A azida sódica pode reagir com os encanamentos e ralos de chumbo ou cobre, originando azidas metálicas altamente explosivas. Ao descartar os restos de reativos, fazê-lo em abundante volume de água. Todos os materiais de origem humana utilizados na preparação deste produto foram testados e apresentaram resultados negativos em relação à presença de anticorpos contra os vírus HIV-1/HIV-2 e HCV, assim como para o antígeno de superfície da hepatite B, utilizando um método aprovado pela Food and Drug Administration (USA). ATENÇÃO: MATERIAL DE RISCO BIOLÓGICO. Dado que nenhum método pode oferecer a total segurança da ausência de agentes infecciosos, este produto deve ser manipulado com precaução. Não utilizar os dispositivos se a embalagem de alumínio sofreu danos durante sua conservação. Para evitar erros de contaminação, deve-se utilizar uma nova pipeta para cada teste. Descartar todos os materiais usados em recipientes adequados para material bio-contaminante. 21 Campioni di sangue totale Test comparativo Totale campioni biorapid MONONUCLEOSIS biorapid MONONUCLEOSIS + - + 15 0 - 0 50 Inmunoensayo de un solo paso para la detección de anticuerpos heterófilos de mononucleosis infecciosa en suero, plasma o sangre total. Sumario biorapid MONONUCLEOSIS + Test comparativo - + 142 0 - 2 478 Quando è stato confrontato con un test immunocromatografico (ICT) commerciale, il biorapid MONONUCLEOSIS ha dimostrato una sensibilità del 100% (142/142) e una specificità del 99,6% (478/480). La concordanza totale tra i due test è stata del 99,7%. Quando è stato confrontato con un test commerciale di agglutinazione di particelle di lattice, il biorapid MONONUCLEOSIS ha dimostrato una sensibilità del 100% (120/120) e una specificità del 97,9% (428/437). La concordanza totale tra i due test è stata del 98,4%. Interferenze Per studiare eventuali interferenze, sono stati analizzati campioni con rischio potenziale di dare falsi risultati positivi, includendo 10 campioni di sieri positivi per gli anticorpi IgM anti-toxoplasma, 10 positivi per gli anticorpi IgM anti-CMV e 10 positivi per il fattore reumatoide. Tutti i campioni hanno dato risultati negativo. Riproducibilità Della valutazione della riproducibilità del test biorapid MONONUCLEOSIS si sono incaricati 3 laboratori esterni in cui il test è stato realizzato da personale con un diverso livello di addestramento. In ogni centro, il test è stato eseguito con campioni etichettati aleatoriamente, 3 positivi e 3 negativi, per 3 giorni e tre volte. I risultati ottenuti presentavano una concordanza del 100% con i risultati previsti. Bibliografia Vedere “References” nel testo inglese. La mononucleosis infecciosa (MI) es una enfermedad infecciosa aguda causada por el virus de Epstein-Barr (EBV). In 1932, Paul y Bunnell observaron que los sueros de pacientes que padecían MI contenían anticuerpos heterófilos contra hematíes de cordero. Más tarde fueron también descritas aglutininas contra eritrocitos de otros mamíferos. Las proteínas responsables de esta aglutinación son glicoproteínas de membrana de los glóbulos rojos denominadas antígeno de Paul-Bunnell por varios autores. Estudios realizados muestran que el antígeno de Paul-Bunnell purificado de eritrocitos bovinos es más sensible a los anticuerpos heterófilos de MI que el procedente de otros mamíferos. También se pueden detectar anticuerpos heterófilos contra eritrocitos de cordero, diferentes de los de MI, en sueros de personas sanas, en individuos que han recibido inyecciones de suero y otros. Tradicionalmente los anticuerpos heterófilos de MI se han diferenciado de los otros tipos mediante una prueba de absorción “diferencial” consistente en la observación del distinto comportamiento de los mismos frente a los absorbentes de riñón de cobaya o de glóbulos de buey. El uso del antígeno de Paul-Bunnell altamente purificado en este test, proporciona un método más simple y más sensible para la detección de los anticuerpos heterófilos de MI. Principio biorapid MONONUCLEOSIS es un test inmunocromatográfico de un solo paso. El dispositivo incluye un pocillo para la adición de la muestra (S), un conjugado de partículas de oro coloidal sensibilizadas con antígeno de Paul-Bunnell altamente purificado, y una membrana cromatográfica en la que se ha inmobilizado antígeno de Paul-Bunnell que actuará como captura. La muestra se dosifica en el pocillo correspondiente (S) y empieza a migrar a lo largo de la membrana adjunta. Si la muestra contiene anticuerpos heterófilos de MI, éstos se unirán al conjugado formando un inmunocomplejo. El complejo continúa su migración a lo largo de la membrana donde será capturado por el antígeno de Paul-Bunnell inmobilizado en la zona test (T), dando lugar a la formación de una línea coloreada. El conjugado sobrante se unirá a la zona control (C) formando una segunda línea coloreada, lo que indica el buen funcionamiento del test. Si la muestra contiene anticuerpos heterófilos de MI, en la ventana de lectura aparecerán dos líneas coloreadas indicando un resultado positivo. Si la muestra no contiene anticuerpos heterófilos de MI, sólo aparecerá la línea Control indicando un resultado negativo. Si no aparece ninguna línea o si sólo aparece la línea Test, el resultado no es válido. Componentes REF 3000-1027. - TEST 25 dispositivos biorapid MONONUCLEOSIS. - DIL 1 x 2 ml tampón diluyente. Tampón Tris con aditivos. - CONTROL + 1 x 1 ml control positivo. IgG de conejo contra el antígeno de Paul-Bunnell diluido en un tampón. Contiene azida sódica < 0,1%. - CONTROL - 1 x 1 ml control negativo. Suero humano negativo diluido. Contiene azida sódica < 0,1%. - PIPETTES 25 pipetas desechables. Precauciones biorapid MONONUCLEOSIS es sólo para el diagnóstico IN VITRO. La azida sódica puede reaccionar con tuberías y desagües de plomo o cobre dando lugar a azidas metálicas altamente explosivas. Al desechar los restos de reactivos, deje correr agua abundante. Todo el material de origen humano utilizado en la preparación de este producto ha sido encontrado negativo a la presencia de anticuerpos de los virus HIV-1/HIV-2 y HCV, así como a la del antígeno de superficie de la hepatitis B, utilizando un método aprobado por la Food and Drug Administration (USA). ATENCIÓN: MATERIAL DE RIESGO BIOLÓGICO. Ya que ningún método puede ofrecer la total seguridad de la ausencia de agentes infecciosos, este producto debe manejarse con precaución. No utilizar los dispositivos si la bolsa de aluminio ha sido dañada durante su conservación. Para evitar errores de contaminación, debe utilizarse una nueva pipeta en cada ensayo. Depositar todos los materiales usados en recipientes adecuados para material biocontaminante. 20 5 Conservación Los reactivos permanecerán inalterados hasta la fecha de caducidad indicada en la etiqueta, si se conservan entre 2-8°C. No congelar. Los dispositivos y el tampón diluyente pueden conservarse por separado entre 2 y 25°C. biokit biokit biokit Recolección de la muestra SUERO o PLASMA: Usar suero fresco o plasma (EDTA, heparina, citrato). Otros anticoagulantes deben ser comprobados antes de utilizarse. Las muestras pueden ser conservadas durante 2 días entre 2 y 8°C. Si es por un período de tiempo más largo las muestras deben ser congeladas (-20°C). Evitar congelar y descongelar las muestras repetidamente. Homogeneizar las muestras antes de analizarlas. No se deben utilizar muestras hemolizadas, turbias, o lipémicas. Las muestras que contienen material particulado pueden dar resultados inconsistentes y deben clarificarse por centrifugación antes de realizar el ensayo. C C C T T T S S S SANGRE TOTAL: Recoger la sangre en tubos con anticoagulante (EDTA, heparina, citrato). Otros anticoagulantes deben ser comprobados antes de utilizarse. Las muestras de sangre total pueden ser conservadas durante 2 días entre 2 y 8°C. POSITIVO Control de calidad Limitazioni della procedura La línea azul que se observa en la zona control (C) es un control de fabricación y no interfiere en el funcionamiento del test. Cualquier resto de colorante azul que pudiera verse en la ventana en el momento de realizar la lectura, no interfiere en los resultados. - NEGATIVO NON VALIDO I risultati vanno interpretati tenendo presente le informazioni cliniche, ematologiche e sierologiche del paziente. In alcuni casi, in pazienti sintomatici della MI gli anticorpi eterofili raggiungono tardi livelli rilevabili. Se i sintomi persistono, è consigliabile ripetere il test dopo un paio di giorni. Alcuni pazienti, soprattutto i bambini e gli adolescenti, possono rimanere sempre negativi. Secondo quanto pubblicato, solo un 80-90% degli adulti e meno del 50% dei ragazzi sviluppano anticorpi eterofili. In alcuni individui, livelli rilevabili di anticorpi eterofili possono persistere mesi e, più raramente, anni. La línea Control (C) es un control interno del procedimiento. Si el test ha sido realizado correctamente y el dispositivo funciona debidamente, esta línea siempre debe aparecer. - Para comprobar el correcto funcionamiento de biorapid MONONUCLEOSIS utilizar los controles incluidos en el kit. Si no se obtienen los resultados esperados, no utilizar el kit. Valori attesi Procedimiento Vari studi su donatori di sangue indicano che gli anticorpi eterofili della MI sono presenti nello 0,9-1,7% della popolazione. Dato che la presenza di tali anticorpi indica un'infezione abbastanza recente, questi risultati suggeriscono che l'effettiva incidenza della malattia potrebbe superare il numero dei casi diagnosticati. PARA SUERO, PLASMA y CONTROLES: - Dejar que los dispositivos y las muestras alcancen la temperatura ambiente antes de utilizarlos. Extraer el dispositivo de la bolsa y colocarlo sobre una superficie plana. Rotular el dispositivo con el nombre del paciente o número de identificación. Mantener la pipeta en posición vertical y dosificar 3 gotas (75 µl) de suero o plasma en el pocillo de muestra (S). Para los controles positivo y negativo, mantener el gotero en posición vertical y dosificar 2 gotas en el pocillo de muestra (S). Immediatamente después, añadir 2 gotas de tampón diluyente, manteniendo el gotero en posición vertical. Leer el resultado a los 5 minutos, luego desechar el dispositivo. No leer el resultado pasados los 5 minutos. Un totale di 622 campioni (296 sieri, 261 plasmi e 65 campioni di sangue totale) sono stati valutati internamente ed esternamente utilizzando il biorapid MONONUCLEOSIS e un altro test immunocromatografico (ICT) commerciale. Tutti i campioni sono stati confermati positivi o negativi con un test commerciale di agglutinazione di particelle di lattice. Risultati Campioni di siero PARA SANGRE TOTAL: - Caratteristiche di funzionalità Dejar que los dispositivos y las muestras alcancen la temperatura ambiente antes de utilizarlos. Extraer el dispositivo de la bolsa y colocarlo sobre una superficie plana. Rotular el dispositivo con el nombre del paciente o número de identificación. Mantener la pipeta en posición vertical y dosificar 2 gotas (50 µl) de sangre en el pocillo de muestra (S). Inmediatamente después, añadir 3 gotas de tampón diluyente, manteniendo el gotero en posición vertical. Leer el resultado a los 5 minutos, luego desechar el dispositivo. No leer el resultado pasados los 5 minutos. biorapid MONONUCLEOSIS Test comparativo Interpretación de resultados + - + 75 0 - 1* 220 (*) il campione discrepante ha dato un risultato negativo con un test di agglutinazione di lattice. POSITIVO Si son visibles dos líneas coloreadas, Test y Control, el resultado es positivo. Cualquier color en la línea Test (T) debe ser interpretado como resultado positivo incluso si el color es menos intenso que el de la línea Control (C). Campioni di plasma biorapid MONONUCLEOSIS NEGATIVO Si sólo es visible la línea Control (C), el resultado es negativo. Test comparativo NO VÁLIDO Si no aparece ninguna línea coloreada o sólo aparece la línea Test, el resultado no es válido. La muestra debe repetirse utilizando otro dispositivo. + - + 52 0 - 1* 208 (*) il campione discrepante ha dato un risultato negativo con un test di agglutinazione di lattice. 6 19 Conservazione I reagenti, se conservati a 2-8°C, sono stabili fino alla data di scadenza indicata sull'etichetta. Non congelare. I dispositivi e il tampone diluente possono essere conservati a parte a 2-25°C. biokit biokit biokit Raccolta dei campioni SIERO OU PLASMA: Usare siero fresco o plasma (EDTA, eparina, citrato). Altri anticoagulanti dovranno essere valutati prima dell’uso. I campioni possono essere conservati a 2-8°C per non più di 2 giorni. Per tempi più lunghi devono essere congelati (-20°C). Non procedere a ripetuti congelamenti e scongelamenti. Miscelare i campioni prima dell’analisi. Non utilizzare campioni emolizzati, torbidi o lipemici. I campioni contenenti materiale particolato possono dare risultati contraddittori e devono essere chiarificati mediante centrifuga prima del test. SANGUE TOTALE: C C C T T T S S S Raccogliere il sangue in provette con anticoagulante (EDTA, eparina, citrato). Gli altri anticoagulanti devono essere valutati prima dell'uso. I campioni di sangue totale possono essere conservati per 2 giorni a 2-8°C. Controllo di qualità POSITIVO La striscia azzurra osservabile nella zona di controllo (C) è un controllo di fabbrica e non interferisce nel funzionamento del test. Qualunque resto di colorante azzurro eventualmente presente sulla finestrella durante la lettura non influenza i risultati. La striscia Controllo (C) è un controllo interno del procedimento e apparirà tutte le volte che il test sia stato eseguito correttamente e il dispositivo funzioni a dovere. - - PER SIERO, PLASMA E CONTROLLI: - Attendere che i dispositivi e i campioni raggiungano la temperatura ambiente prima di utilizzarli. Estrarre il dispositivo dalla busta e sistemarlo su una superficie piana. Apporre una etichetta sul dispositivo con il nome del paziente o un numero di identificazione. Tenere la pipetta in posizione perpendicolare e dispensare 3 gocce (75 µl) di siero o plasma nel pozzetto per il campione (S). Per i controlli positivo e negativo mantenere il contagocce in posizione perpendicolare e dispensare 2 gocce nel pozzetto per il campione (S). Subito dopo, aggiungere 2 gocce di tampone diluente, tenendo il contagocce in posizione perpendicolare. Dopo 5 minuti leggere il risultato e gettare il dispositivo. Non leggere nessun risultato una volta trascorsi i 5 minuti. PER SANGUE TOTALE: - Attendere che i dispositivi e i campioni raggiungano la temperatura ambiente prima di utilizzarli. Estrarre il dispositivo dalla busta e sistemarlo su una superficie piana. Apporre una etichetta sul dispositivo con il nome del paziente o un numero di identificazione. Tenere la pipetta in posizione perpendicolare e dispensare 2 gocce (50 µl) di sangue nel pozzetto per il campione (S). Subito dopo, aggiungere 3 gocce di tampone diluente, tenendo il contagocce in posizione perpendicolare. Dopo 5 minuti leggere il risultato e gettare il dispositivo. Non leggere nessun risultato una volta trascorsi i 5 minuti. Los resultados deben ser interpretados teniendo en consideración la información clínica, hematológica y serológica del paciente. Ocasionalmente niveles detectables de anticuerpos heterófilos se desarrollan tardíamente en pacientes sintomáticos de MI. Si los síntomas persisten se recomienda repetir el ensayo al cabo de unos días. Algunos pacientes, especialmente niños y adolescentes, pueden persistir permanentemente negativos. Se ha publicado que sólo de un 80 a un 90% de los adultos y menos de un 50% de los jóvenes desarrollan anticuerpos heterófilos. En algunos individuos pueden persistir niveles detectables de anticuerpos heterófilos durante meses y, más raramente, durante años. Valores previstos Diferentes estudios realizados en donantes de sangre indican que la presencia de anticuerpos heterófilos de MI oscila entre un 0,9 y un 1,7% de la población. Puesto que la presencia de dichos anticuerpos indica una infección relativamente reciente, estos resultados sugieren que la incidencia real de la enfermedad puede ser mayor que el número de casos diagnosticados. Características funcionales Un total de 622 muestras (296 sueros, 261 plasmas y 65 sangre total) fueron evaluadas internamente y en una evaluación externa utilizando el test biorapid MONONUCLEOSIS y otro test inmunocromatográfico (ICT) comercial. Todas las muestras fueron confirmadas como positivas o negativas mediante un test comercial de aglutinación de partículas de látex. Resultados Muestras de suero Interpretazione dei risultati biorapid MONONUCLEOSIS Test comparativo POSITIVO Se le due strisce colorate (Test e Controllo) sono visibili, il risultato è positivo. Qualsiasi colore nella striscia Test (T) va interpretato come risultato positivo, anche se il colore è meno intenso di quello della striscia Controllo (C). NEGATIVO + - + 75 0 - 1* 220 (*) La muestra discrepante dio un resultado negativo con un test de aglutinación de látex. Muestras de plasma Se appare colorata solo la striscia Controllo (C), il risultato è negativo. biorapid MONONUCLEOSIS NON VALIDO Se non compare nessuna striscia colorata, o se appare solo la striscia Test, il risultato non è valido. Ripetere utilizzando un altro dispositivo. NO VÁLIDO Limitaciones del procedimiento Per verificare il corretto funzionamento del biorapid MONONUCLEOSIS utilizzare i controlli inclusi nel kit. Se i risultati non sono quelli previsti, non utilizzare il kit. Procedura NEGATIVO Test comparativo + - + 52 0 - 1* 208 (*) La muestra discrepante dio un resultado negativo con un test de aglutinación de látex. 18 7 Muestras de sangre total Test comparativo Total muestras biorapid MONONUCLEOSIS biorapid MONONUCLEOSIS + - + 15 0 - 0 50 Immunotest a un solo passaggio per la determinazione di anticorpi eterofili di mononucleosi infettiva in siero, plasma o sangue totale. biorapid MONONUCLEOSIS Test comparativo + - + 142 0 - 2 478 Sommario Cuando se comparó con un test inmunocromatográfico (ICT) comercial, biorapid MONONUCLEOSIS mostró una sensibilidad del 100% (142/142) y una especificidad del 99,6% (478/480). La concordancia total entre los dos tests fue del 99,7%. Cuando se comparó con un test commercial de aglutinación de partículas de látex, el test biorapid MONONUCLEOSIS demostró una sensibilidad del 100% (120/120) y una especificidad del 97,9% (428/437). La concordancia total entre los dos tests fue del 98,4%. Interferencias Para estudiar posibles interferencias, se analizaron muestras con riesgo potencial de producir resultados falsos positivos. Se incluyeron 10 muestras de suero positivas para anticuerpos IgM anti-toxoplasma, 10 positivas para anticuerpos IgM anti-CMV y 10 positivas para factor reumatoide. Todas las muestras dieron resultado negativo. Reproducibilidad Una evaluación de la reproducibilidad del test biorapid MONONUCLEOSIS fue llevada a cabo en 3 laboratorios externos donde la prueba fue realizada por personal de distinto nivel de entrenamiento. En cada centro, la prueba se realizó con muestras rotuladas al azar, 3 positivas y 3 negativas, durante 3 días y por triplicado. Los resultados obtenidos mostraron un 100% de concordancia con los resultados esperados. Bibliografía Ver “References” del texto inglés. La mononucleosi infettiva (MI) è una malattia infettiva acuta causata dal virus di Epstein-Barr (EBV). Nel 1932, Paul e Bunnell avevano osservato che i sieri di pazienti affetti da MI contenevano anticorpi eterofili anti-emazie di pecora, ma più tardi sono state descritte anche agglutinine anti-eritrociti di altri mammiferi. Le proteine responsabili di questa agglutinazione sono le glicoproteine di membrana dei globuli rossi, denominate da vari autori antigene di Paul-Bunnell. Alcuni studi mostrano che l’antigene Paul-Bunnell purificato dagli eritrocici bovini è più sensibile agli anticorpi eterofili di MI di quello di altri mammiferi. Anticorpi eterofili anti-eritrociti di pecora, distinti da quelli della MI, sono riscontrabili anche nei sieri di individui sani, di persone che hanno ricevuto iniezioni di siero e di altri soggetti. Tradizionalmente, per distinguere gli anticorpi eterofili della MI dagli altri tipi, si è utilizzato un test di assorbimento “differenziale” con emazie bovine e tessuto renale di cavia. In questo test, l’uso dell’antigene di Paul-Bunnell altamente purificato offre un metodo più semplice e sensibile per la determinazione degli anticorpi eterofili di MI. Principio Il biorapid MONONUCLEOSIS è un test immunocromatografico a un solo passaggio. Il dispositivo comprende un pozzetto per il campione (S), un coniugato di particelle di oro colloidale sensibilizzate con antigene di Paul-Bunnell altamente purificato, e una membrana cromatografica su cui è stato immobilizzato antigene di Paul-Bunnell che agirà come elemento di cattura. Il campione viene dispensato nel relativo pozzetto (S) e inizia a migrare lungo la membrana annessa. Se il campione contiene anticorpi eterofili di MI, questi si uniranno al coniugato formando un immunocomplesso che proseguirà la propria migrazione lungo la membrana dove sarà catturato dall’antigene di Paul-Bunnell immobilizzato nella zona del test (T), provocando la formazione di una striscia colorata. Il coniugato che avanza verrà unito nella zona di controllo (C) formando una seconda striscia colorata, a indicazione del corretto funzionamento del test. Se il campione contiene anticorpi eterofili di MI, sulla finestrella di lettura appariranno due strisce colorate indicanti un risultato positivo. Se il campione non contiene anticorpi eterofili di MI, apparirà solo la striscia Controllo indicante un risultato negativo. Se non appare nessuna striscia o se appare solo la striscia Test, il risultato non è valido. Componenti REF 3000-1027. - TEST 25 dispositivi biorapid MONONUCLEOSIS. - DIL 1 x 2 ml tampone diluente. Tampone Tris con additivi. - CONTROL + 1 x 1 ml controllo positivo. IgG di coniglio contro l’antigene di Paul-Bunnell diluito in un tampone. Contiene sodio azide < 0,1%. - CONTROL - 1 x 1 ml controllo negativo. Siero umano negativo diluito. Contiene sodio azide < 0,1%. - PIPETTES 25 pipette monouso. Precauzioni biorapid MONONUCLEOSIS è per uso diagnostico IN VITRO. La sodio azide può reagire a contatto con tubature e scarichi in piombo o in rame, formando azidi metalliche altamente esplosive. Far scorrere acqua in abbondanza quando si gettano i residui dei reagenti. Tutto il materiale di origine umana utilizzato nella preparazione di questo prodotto è risultato negativo per anticorpi dei virus HIV-1/HIV-2 e HCV come pure per l’antigene di superficie dell’epatite B, usando metodi approvati dalla Food and Drug Administration (USA). ATTENZIONE: MATERIALE POTENZIALMENTE BIOPERICOLOSO. Poiché nessun metodo può offrire la totale sicurezza dell’assenza di agenti infettivi, questo prodotto deve essere manipolato con precauzione. Non utilizzare i dispositivi se la busta di alluminio è stata danneggiata nel corso della conservazione. Per evitare errori dovuti a contaminazione utilizzare una pipetta nuova per ogni test. Depositare tutti i materiali utilizzati in recipienti idonei per materiale biocontaminante. 8 17 Échantillons de sang total Test comparatif biorapid MONONUCLEOSIS biorapid MONONUCLEOSIS + - + 15 0 - 0 50 Einschritt-Immuntest zur Bestimmung heterophiler IM-Antikörper infektiöser Mononukleose in Serum, Plasma oder Vollblut. bei Zusammenfassung Total échantillons biorapid MONONUCLEOSIS + Test comparatif - + 142 0 - 2 478 Comparé à un test immunochromatographique (ICT) commercial, biorapid MONONUCLEOSIS a montré une sensibilité de 100% (142/142) et une spécificité de 99,6% (478/480). La concordance totale entre les deux tests a été de 99,7%. Comparé à un test commercial d’agglutination de particules de latex, le test biorapid MONONUCLEOSIS a montré une sensibilité de 100% (120/120) et une spécificité de 97,9% (428/437). La concordance totale entre les deux tests a été de 98,4%. Interférences Pour étudier d’éventuelles interférences, des échantillons présentant un risque potentiel de produire des résultats faux positifs ont été testés. Ce test comprenait 10 échantillons de sérum positifs pour les anticorps IgM anti-toxoplasma, 10 positifs pour les anticorps IgM anti-CMV et 10 positifs pour le facteur rhumatoïde. Tous les échantillons ont donné des résultats négatifs. Reproductibilité Une étude de reproductibilité du test biorapid MONONUCLEOSIS a été réalisée dans 3 laboratoires externes par des laborantins qui avaient différents niveaux d’expérience. Dans chaque centre, le test a porté sur des échantillons identifiés au hasard, 3 positifs et 3 négatifs, pendant 3 jours et il a été répété 3 fois. Les résultats obtenus ont donné 100% de concordance avec les résultats attendus. Bibliographie Voir “Références” du texte anglais. Die infektiöse Mononukleose (IM) ist eine schwere Infektionskrankheit, die von dem Epstein-Barr-Virus (EBV) ausgelöst wird. Paul und Bunnell beobachteten im Jahr 1932, dass das Serum von IM-Patienten heterophile Antikörper gegen Schafserythrozyten enthielt. Später wurden auch Agglutinine gegen Erythrozyten anderer Säugetiere beschrieben. Die für diese Agglutination verantwortlichen Proteine sind Glykoproteine der Erythrozytenmembranen, die von verschiedenen Autoren als Paul-Bunnell-Antigene bezeichnet werden. In Studien wurde nachgewiesen, dass das aus roten Blutkörperchen von Rinderzellen gereinigte Paul-Bunnell-Antigen sensitiver auf die heterophilen IM-Antikörper reagiert als das Antigen von anderen Säugetieren. Heterophile Antikörper gegen Schafserythrozyten, die sich von den IM-Antikörpern unterscheiden, können auch in Seren von gesunden Personen, von Personen, die Seruminjektionen erhalten haben, und anderen beobachtet werden. Früher wurden die heterophilen IM-Antikörper von anderen heterophilen Antikörpern durch einen “Differentialabsorptionstest” mit Nierengewebe von Versuchskaninchen und Rinder-Erythrozyten unterschieden. Die Verwendung des hochreinen Paul-Bunnell-Antigens in diesem Test ist eine einfachere und sensitivere Methode zur Bestimmung der heterophilen IM-Antikörper. Prinzip biorapid MONONUCLEOSIS ist ein immunchromatographischer Einschritt-Test. Das Gerät enthält eine Vertiefung für die Zugabe der Probe (S), ein Konjugat aus Kolloidalgold-Partikeln, die mit hochreinem Paul-Bunnell-Antigen beschichtet sind, und eine chromatographische Membran, auf der Paul-Bunnell-Antigen immobilisiert ist, welches als Fangantikörper wirkt. Die Probe wird in der entsprechenden Vertiefung (S) aufgetragen und beginnt auf der benachbarten Membran entlang zu migrieren. Wenn die Probe heterophile IM-Antikörper enthält, werden sich diese an das Konjugat binden und ein Immunkomplex bilden. Der Komplex setzt seine Migration auf der Membran fort, wo es von dem in der Testzone (T) immobilisierten Paul-Bunnell-Antigen gefangen wird, was zur Entstehung einer Farblinie führt. Das restliche Konjugat wird sich an die Kontrollzone (C) binden und dabei eine weitere Farblinie bilden, die den korrekten Ablauf des Tests anzeigt. Wenn die Probe heterophile IM-Antikörper enthält, erscheinen im Sichtfenster zwei Farblinien, die ein positives Ergebnis anzeigen. Wenn die Probe keine heterophilen IM-Antikörper enthält, erscheint nur die Kontrolllinie, die ein negatives Ergebnis bedeutet. Wenn keine Linie erscheint oder nur die Testlinie zu sehen ist, ist das Ergebnis ungültig. Bestandteile REF 3000-1027. - TEST 25 Geräte biorapid MONONUCLEOSIS. - DIL 1 x 2 ml Lösungsmittel-Puffer. Tris-Puffer mit Zusatzmitteln. - CONTROL + 1 x 1 ml positive Kontrolle. Kaninchen-IgG gegen Paul-Bunnell-Antigen verdünnt in Pufferlösung. Enthält < 0,1% Natriumazid. - CONTROL - 1 x 1 ml negative Kontrolle. Verdünntes, negatives Humanserum. Enthält < 0,1% Natriumazid. - PIPETTES 25 Einwegpipetten. Vorsichtsmaßnahmen biorapid MONONUCLEOSIS ist ausschliesslich für die IN VITRO-Diagnostik bestimmt. Natriumazid kann auf Blei- oder Kupfer-Rohrleitungen und -Abflüsse reagieren und zu hochexplosiven Metallaziden führen. Beim Entfernen der Reagenzreste muss mit genügend Wasser nachgespült werden. Das gesamte zur Herstellung dieses Produkts verwendete Humanmaterial ist nach den Richtlinien der FDA (USA) auf die Anwesenheit von HBsAg, HIV-1/HIV-2 und HCV Antikörpern getestet und für negativ befunden worden. ACHTUNG: BIOLOGISCH GEFÄHRLICHES MATERIAL. Da kein Nachweisverfahren die Abwesenheit von infektiösen Agenzien garantieren kann, ist dieses Produkt mit entsprechenden Vorsichtsmassnahmen zu handhaben. Die Geräte dürfen nicht verwendet werden, wenn der Aluminiumbeutel während der Lagerung beschädigt worden ist. Um Fehler durch Kontamination zu vermeiden, muss bei jedem Test neue Pipette verwendet werden. Alle benutzten Materialien sind separat in Behältern für biologische Abfälle zu entsorgen. 16 9 Lagerung Bei einer Lagerungstemperatur von 2-8°C bleiben die Reagenzien bis zum auf dem Etikett angegebenen Verfallsdatum haltbar. Nicht einfrieren. Die Geräte und der Lösungsmittel-Puffer können getrennt voneinander bei 2 bis 25°C gelagert werden. biokit biokit biokit Probenmaterial SERUM ODER PLASMA: Frisches Serum oder Plasma (EDTA, Heparin, Zitrat) benutzen. Andere Antikoagulantien müssen zuvor evaluiert werden. Die Seren sind bei 2-8 °C zu lagern. Bei Lagerung über 2 Tagen sollten die Proben bei (-20°C) eingefroren werden. Mehrmaliges Einfrieren und Auftauen vermeiden. Vor Testdurchführung Proben homogenisieren. Es dürfen keine hämolisierten, trüben oder lipämischen Proben verwendet werden. Die Proben, die Partikel enthalten, können zu inkonsistenten Ergebnissen führen und müssen vor dem Test durch Zentrifugation geklärt werden. C C C T T T S S S VOLLBLUT: Das Blut wird in Rohre mit Antikoagulant (EDTA, Heparin, Zitrat) gefüllt. Andere Antikoagulantien müssen vor der Verwendung geprüft werden. Die Vollblutproben können 2 Tage bei 2 bis 8°C gelagert werden. POSITIF Qualitätskontrolle Die blaue Linie, die in der Kontrollzone (C) erscheint, ist eine Herstellungskontrolle und hat keinen Einfluß auf den Testablauf. Sollten beim Ablesen Reste vom blauen Farbstoff im Sichtfenster zu erkennen sein, so haben diese keinerlei Einfluß auf die Testergebnisse. Bei der Kontrolllinie (C) handelt es sich um eine interne Ablaufkontrolle. Diese Linie muss immer dann erscheinen, wenn der Test korrekt durchgeführt wurde und das Gerät ordnungsgemäß funktioniert. Zur Überprüfung des korrekten Funktionierens des biorapid MONONUCLEOSIS werden die im Kit enthaltenen Kontrollen verwendet. Wenn die erhaltenen Werte von den erwarteten abweichen, darf das Kit nicht verwendet werden. Durchführung NÉGATIF NON VALABLE Limites de la procédure - - Les résultats seront interprétés en accord avec le bilan clinique, hématologique et sérologique du patient. Occasionnellement, chez des patients symptomatiques de la MI, les concentrations détectables d’anticorps hétérophiles apparaissent tardivement. Si les symptômes persistent, il est recommandé de réaliser un nouveau test quelques jours après. Certains patients ne développent pas d’anticorps hétérophiles, en particulier les enfants et les adolescents. D’après la littérature, 80 à 90% des adultes et moins de 50% des enfants développent des anticorps hétérophiles. Chez certains sujets, des concentrations détectables d’anticorps hétérophiles peuvent persister pendant des mois et plus rarement pendant des années. FÜR SERUM, PLASMA UND KONTROLLEN: Valeurs attendues - Plusieurs études sur la présence d’anticorps hétérophiles de la MI chez les donneurs de sang rapportent son incidence sur 0,9 à 1,7% de la population. Puisque la présence de ces anticorps indique une infection relativement récente, ces résultats suggèrent que l’incidence réelle de la maladie est plus élevée que le nombre de cas diagnostiqués. - Die Geräte und Proben vor der Anwendung auf Raumtemperatur bringen. Das Gerät aus der Beutel entnehmen und auf eine flache Oberfläche stellen. Das Gerät mit dem Namen oder der Kennnummer des Patienten beschriften. Die Pipette senkrecht halten und 3 Tropfen (75 µl) Serum oder Plasma auf die Probenvertiefung (S) geben. Für die positiven und negativen Kontrollen die Tropfflasche senkrecht halten und 2 Tropfen auf die Probenvertiefung (S) geben. Gleich danach 2 Tropfen Lösungsmittel-Puffer aus der senkrecht gehaltenen Tropfflasche hinzufügen. Das Ergebnis nach 5 Minuten ablesen, danach das Gerät entsorgen. Das Ergebnis nicht mehr ablesen, wenn mehr als 5 Minuten vergangen sind. FÜR VOLLBLUT: Die Geräte und Proben vor der Anwendung auf Raumtemperatur bringen. Das Gerät aus der Beutel entnehmen und auf eine flache Oberfläche stellen. Das Gerät mit dem Namen oder der Kennnummer des Patienten beschriften. Die Pipette senkrecht halten und 2 Tropfen (50 µl) Blut auf die Probenvertiefung (S) geben. Gleich danach 3 Tropfen Lösungsmittel-Puffer aus der senkrecht gehaltenen Tropfflasche hinzufügen. Das Ergebnis nach 5 Minuten ablesen, danach das Gerät entsorgen. Das Ergebnis nicht mehr ablesen, wenn mehr als 5 Minuten vergangen sind. Caractéristiques fonctionnelles Un total de 622 échantillons (296 sérums, 261 plasmas et 65 sang total) ont été soumis à une analyse interne et à une analyse externe en utilisant le test biorapid MONONUCLEOSIS et un autre test immunochromatographique (ICT) commercial. Les résultats, positif ou négatif, de tous les échantillons ont été confirmés par un test commercial d’agglutination de particules de latex. Résultats Échantillons de sérum biorapid MONONUCLEOSIS Test comparatif Auswertung der Ergebnisse POSITIV Wenn zwei Farblinien, Test und Kontrolle, sichtbar sind, ist das Ergebnis positiv. Jede in der Testlinie (T) sichtbare Färbung ist als positives Ergebnis zu interpretieren, auch wenn deren Farbintensität geringer als die der Kontrolllinie (C) ist. Wenn nur die Kontrolllinie (C) sichtbar ist, ist das Ergebnis negativ. UNGÜLTIG Wenn keine Farblinie erscheint oder nur die Testlinie zu erkennen ist, ist das Ergebnis ungültig. Die Probe muss mit einem anderen Gerät wiederholt werden. - 75 0 - 1* 220 (*) L’échantillon discordant a donné un résultat négatif au test d’agglutination de latex. Échantillons de plasma NEGATIV + + Test comparatif biorapid MONONUCLEOSIS + - + 52 0 - 1* 208 (*) L’échantillon discordant a donné un résultat négatif au test d’agglutination de latex. 10 15 Conservation Les réactifs resteront stables jusqu’à la date de péremption indiquée sur l’étiquette, à condition d’être conservés entre 2-8°C. Ne pas congeler. Les dispositifs et le tampon diluant peuvent être conservés à part entre 2 et 25°C. biokit biokit biokit Prélèvement de l’échantillon SÉRUM OU PLASMA: Utiliser du sérum ou du plasma frais (EDTA, héparine, citrate). D’autres anticoagulants doivent être évalués avant leur utilisation. Les échantillons peuvent être conservés à 2-8°C pendant 2 jours. Pour des périodes plus longues, les échantillons doivent être congelés (-20°C). Éviter les cycles répétés de congélation/décongélation des échantillons. Bien mélanger les échantillons avant analyse. Ne pas utiliser d’échantillons hémolysés, troubles ou lipémiques. Les échantillons contenant du matériel en particules peuvent donner des résultats inconsistants et doivent être éclaircis par centrifugation avant le test. C C C T T T S S S SANG TOTAL: Prélever le sang dans des tubes avec de l’anticoagulant (EDTA, héparine, citrate). D’autres anticoagulants demandent à être vérifiés avant utilisation. Les échantillons de sang total peuvent être conservés pendant 2 jours entre 2 et 8°C. Contrôle de qualité POSITIV UNGÜLTIG Begrenzung der Methode - La ligne bleue qui apparaît dans la zone contrôle (C) correspond à un contrôle de fabrication et n’interfère pas dans le fonctionnement du test. Toute trace de colorant bleu, visible à la lecture de la fenêtre, n’interfère pas sur les résultats. NEGATIV - La ligne Contrôle (C) est un contrôle interne de la procédure. Si le test a été réalisé correctement et si le dispositif fonctionne bien, cette ligne doit obligatoirement apparaître. Die Ergebnisse müssen unter Berücksichtigung aller weiteren klinischen, hämatologischen und serologischen Daten des Patienten ausgewertet werden. Es besteht die Möglichkeit, dass sich feststellbare Titer heterophiler Antikörper erst verzögert in Patienten mit IM-Symptomatik entwickeln. Falls die Symptome bestehen bleiben, sollte der Test nach einigen Tagen wiederholt werden. Einige Patienten, insbesondere Kinder und Jugendliche, zeigen durchgehend negative Ergebnisse. In der Literatur wird berichtet, dass nur 80 bis 90% der Erwachsenen und weniger als 50% der Jugendlichen heterophile Antikörper entwickeln. Feststellbare Titer heterophiler Antikörper können bei einigen Patienten über Monate und, in selteneren Fällen, über Jahre hinweg bestehen bleiben. Pour vérifier le bon fonctionnement du biorapid MONONUCLEOSIS, utiliser les contrôles du coffret. Si les résultats attendus n’apparaissent pas, ne pas utiliser le coffret. - Procédure Erwartete Werte POUR LE SÉRUM, LE PLASMA ET LES CONTRÔLES: Verschiedene Studien zur Anwesenheit heterophiler IM-Antikörper in Blutspendern zeigen eine Krankheitsinzidenz von 0,9 bis 1,7% der Bevölkerung. Die Anwesenheit von heterophilen Antikörpern zeigt eine relativ frische Infektion an, sodass die wirkliche Inzidenz der Krankheit höher als die Anzahl der diagnostizierten Fälle sein kann. - Attendre que les dispositifs et les échantillons soient à température ambiante avant de les utiliser. Retirer le dispositif de la poche et le poser sur une surface plane. Étiqueter le dispositif avec le nom du patient ou le numéro d’identification. Maintenir la pipette en position verticale et doser 3 gouttes (75 µl) de sérum ou de plasma dans le puits échantillon (S). Pour les contrôles positif et négatif, maintenir le compte-gouttes en position verticale et doser 2 gouttes dans le puits échantillon (S). Immédiatement après, ajouter 2 gouttes de tampon diluant, en maintenant le compte-gouttes en position verticale. Lire le résultat au bout de 5 minutes, mais jamais au-delà, et jeter le dispositif. Es wurden insgesamt 622 Proben (296 Seren, 261 Plasmen und 65 Vollblut) sowohl intern als auch in einer externen Prüfung mit dem Test biorapid MONONUCLEOSIS und einem anderen kommerziellen immunchromatographischen Test (ICT) geprüft. Alle Proben wurden mit einem kommerziellen Latexpartikel-Agglutinationstest als positiv bzw. negativ bestätigt. Ergebnisse POUR LE SANG TOTAL: - Charakteristika des Tests Serumproben Attendre que les dispositifs et les échantillons soient à température ambiante avant de les utiliser. Retirer le dispositif de la poche et le poser sur une surface plane. Étiqueter le dispositif avec le nom du patient ou le numéro d’identification. Maintenir la pipette en position verticale et doser 2 gouttes (50 µl) de sang dans le puits échantillon (S). Immédiatement après, ajouter 3 gouttes de tampon diluant, en maintenant le compte-gouttes en position verticale. Lire le résultat au bout de 5 minutes, mais jamais au-delà, et jeter le dispositif. biorapid MONONUCLEOSIS Vergleichstest Interprétation des résultats POSITIF Si deux lignes colorées, Test et Contrôle, apparaissent, le résultat est positif. Toute coloration de la ligne Test (T) doit être interprétée comme un résultat positif, même si la couleur est moins intense que celle de la ligne Contrôle (C). + - + 75 0 - 1* 220 (*) Die abweichende Probe erbrachte ein negatives Ergebnis mit einem Latex-Agglutinationstest. Plasmaproben biorapid MONONUCLEOSIS NÉGATIF Si seule la ligne Contrôle (C) est visible, le résultat est négatif. NON VALABLE Vergleichstest Si aucune ligne colorée n’est visible ou si seule la ligne Test apparaît, le résultat n’est pas valable. Il faut alors recommencer le test en utilisant un autre dispositif. + - + 52 0 - 1* 208 (*) Die abweichende Probe erbrachte ein negatives Ergebnis mit einem Latex-Agglutinationstest. 14 11 Vollblutproben biorapid MONONUCLEOSIS biorapid MONONUCLEOSIS Vergleichstest + - + 15 0 - 0 50 Immunotest en une seule étape pour la détection d’anticorps hétérophiles de la mononucléose infectieuse dans le sérum, le plasma ou le sang total. Sommaire Gesamtproben biorapid MONONUCLEOSIS Vergleichstest + - + 142 0 - 2 478 Im Vergleich mit einem kommerziellen immunchromatographischen Test (ICT) zeigte biorapid MONONUCLEOSIS eine Sensitivität von 100% (142/142) und eine Spezifität von 99,6% (478/480). Die Gesamtübereinstimmung zwischen den beiden Tests betrug 99,7%. Im Vergleich mit einem kommerziellen Latexpartikel-Agglutinationstest zeigte der Test biorapid MONONUCLEOSIS eine Sensitivität von 100% (120/120) und eine Spezifität von 97,9% (428/437). Die Gesamtübereinstimmung zwischen den beiden Tests betrug 98,4%. Interferenzen Um möglicher Interferenzen zu untersuchen, wurden Proben mit einem potentiellen Risiko, Kreuzreaktionen hervorzurufen, analysiert. Diese Proben bestanden aus 10 positiven Seren für IgM-Antikörper gegen Toxoplasma, 10 für IgM-Antikörper gegen CMV und 10 für den rheumatoiden Faktor. Es wurden dabei keine Interferenzen festgestellt. Reproduzierbarkeit La mononucléose infectieuse (MI) est une maladie infectieuse aiguë causée par le virus d’Epstein-Barr (EBV). En 1932, Paul et Bunnell rapportent que le sérum de patients atteints de MI contenait des anticorps hétérophiles contre les hématies de mouton. Des agglutinines anti-érythrocytes d’autres mammifères ont aussi été décrites. Les protéines responsables de cette agglutination sont des glycoprotéines de membrane de globules rouges, appelées antigènes Paul-Bunnell par plusieurs auteurs. Des études ont démontré que l’antigène de Paul-Bunnell purifié d’érythrocytes bovins était plus sensible aux anticorps hétérophiles de la MI que celui d’autres mammifères. Des anticorps hétérophiles anti-érythrocytes de mouton, différents de ceux de la MI, peuvent aussi être détectés dans le sérum de personnes saines, transfusées ou autres. Traditionnellement, les anticorps hétérophiles de la MI se distinguent d’autres anticorps hétérophiles par un test d’absorption “différentielle” avec des globules rouges bovins et du tissu rénal de cobaye. L’utilisation de l’antigène de Paul-Bunnell hautement purifié dans ce test fournit une méthode plus simple et plus sensible pour la détection des anticorps hétérophiles de la MI. Principe biorapid MONONUCLEOSIS est un test immunochromatographique en une seule étape. Le dispositif comprend un puits pour l’addition de l’échantillon (S), un conjugué de particules d’or colloïdal sensibilisées à l’antigène de PaulBunnell hautement purifié et une membrane chromatographique sur laquelle l’antigène de Paul-Bunnell a été immobilisé pour servir de capture. L’échantillon est dosé dans le puits correspondant (S) et commence à migrer le long de la membrane. Si l’échantillon contient des anticorps hétérophiles de la MI, ceux-ci s’associeront au conjugué et formeront un immunocomplexe. Le complexe poursuit sa migration le long de la membrane où il sera capturé par l’antigène de Paul-Bunnell immobilisé dans la zone test (T) et il s’ensuivra la formation d’une ligne colorée. L’excédent de conjugué rejoindra la zone contrôle (C) et formera une deuxième ligne colorée, ce qui indiquera le bon fonctionnement du test. Die Reproduzierbarkeit des Tests biorapid MONONUCLEOSIS wurde in 3 externen Labors von unterschiedlich geschultem Personal getestet. In jedem Labor wurde der Test mit zufällig beschrifteten Proben, 3 positiven und 3 negativen, 3 Tage lang dreifach getestet. Die Ergebnisse zeigten eine 100%ige Übereinstimmung mit den erwarteten Ergebnissen. Si l’échantillon contient des anticorps hétérophiles de la MI, deux lignes colorées apparaîtront dans la fenêtre de lecture, ce qui indiquera un résultat positif. Si l’échantillon ne contient pas d’anticorps hétérophiles de la MI, seule la ligne Contrôle apparaîtra, ce qui indiquera un résultat négatif. Si aucune ligne n’apparaît à la lecture ou seulement la ligne Test, le résultat n’est pas valable. Literatur Composants Siehe “References” im englischen Text. REF 3000-1027. - TEST 25 dispositifs biorapid MONONUCLEOSIS. - DIL 1 x 2 ml tampon diluant. Tampon Tris et additifs. - CONTROL + 1 x 1 ml contrôle positif. IgG de lapin contre l’antigène de Paul-Bunnell dilué dans un tampon. Contient de l’azide sodique < 0,1%. - CONTROL - 1 x 1 ml contrôle négatif. Sérum humain négatif dilué. Contient de l’azide sodique < 0,1%. - PIPETTES 25 pipettes jetables. Précautions biorapid MONONUCLEOSIS est destiné à un usage diagnostique IN VITRO. L’azide sodique peut réagir au contact de tuyauteries et de bouches d’écoulement des eaux en plomb ou en cuivre. Cette réaction donne lieu à des azides métalliques hautement explosives. Lorsque vous jetez les restes de réactifs, veuillez laisser couler l’eau abondamment. Tout le matériel d’origine humaine utilisé dans le cadre de la préparation de ce produit a donné des résultats négatifs quant à la présence d’anticorps aux virus HIV-1/HIV-2 et HCV, de même qu’à celle de l’antigène de surface de l’hépatite B. La méthode utilisée est agréée par la Food and Drug Administration (USA). ATTENTION: MATÉRIEL À RISQUE BIOLOGIQUE. Compte tenu du fait qu’aucune méthode n’est capable d’offrir une garantie absolue de l’absence d’agents infectieux, ce produit doit être manipulé avec précaution. Ne pas utiliser les dispositifs si la poche en aluminium a été endommagée pendant leur conservation. Pour éviter des erreurs de contamination, il faut utiliser une nouvelle pipette à chaque test. Déposer tout le matériel utilisé dans des récipients conçus pour le matériel bio-contaminant. 12 13
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with IM have heterophile antibodies to sheep erythrocytes. Also were described agglutinins to red blood cells from other mammals. The proteins responsible for this agglutination are glycoproteins f...
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