Public Health Practice Program Office, Centers for Disease Control* Laboratory Directors and AIDS Program, Center for Infectious Reported by: Association of State and Territorial Public Health Please note: This guideline document is obsolete and may not reflect current evidence or best practice and likely contains out-of-date information. Interpretation and Use of the Western Blot Assayįor Serodiagnosis of Human Immunodeficiency Virus Type 1 Infections For assistance, please send e-mail to: Type 508 Accommodation and the title of the report in the subject line of e-mail. The result showed 100 % Sensitivity & 100 % Specificity.Persons using assistive technology might not be able to fully access information in this file. Results were compared with those obtained by the source Institutes and the sensitivity & specificity were evaluated. of India, New Delhi was evaluated at Cancer Research Centre-Mumbai. VIROLOGY LABORATORY, CANCER RESEARCH INSTITUTE-MUMBAI: 250 Coded sera originating from five different national institutes (NICB-Delhi, PGIMER-Chandigarh, CMC-Vellore, NARI-Pune, NICED-Kolkata) as sent by Dept.NIMHANS-BANGALORE- NIMHANS BANGALORE: Evaluation report from National HIV Reference Laboratories of Government of India, claiming Sensitivity and Specificity of HIV 1 & 2 Western Blot to be 100 %.IJMM: Evaluation study by Indian journal of Medical Microbiology claimed 100 % Sensitivity & 100 % Specificity of HIV Western Blot.NICD DELHI: Evaluation Report from National Institute of Communicable Diseases claiming 100 % Sensitivity & 100 % Specificity.If HIV specific antibodies are not present, the band pattern does not meet the required criteria. If antibodies to HIV-2 antigens are present, HIV-2 band is also observed along with some of the other bands. If antibodies to HIV-1 antigens are present in the sera, any two ENVELOPE with one or more of the following bands will be seen: p17, p24, p31, gp41, p51/p55, p66, gp120 & gpl60. After washing the unbound conjugate, substrate (BCIP/NBT) is added which results in the staining of bands. Unbound material is washed off and then the strip is incubated with anti-human IgG conjugated to alkaline phosphatase (AP). Antibodies to HIV- 1 & 2 if present, bind to viral antigens located on the strip. To perform the assay, the strip is incubated with the patient serum/plasma diluted in a buffer. The membrane is cut and packaged as strips. They are then transferred from SDS-PAGE gel on to nitrocellulose membrane, which is also impregnated with HIV-2 antigen(gp36) and a control band. Low molecular weight components migrate faster and are found at the bottom of the gel, while high molecular weight proteins remain near the top. SDS denatures viral components and yields proteins which migrate in the gel according to their molecular weight to produce various bands.
The HIV-1 viral antigens are purified and then separated by SDS gel electrophoresis. The HIV 1&2 Western Blot is manufactured from HIV-1 cell line. Convenient pack size: 5 Tests & 25 Tests.Individual test run possible, separate disposable trays for each strip provided.
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