An international collaborative study on the detection of an HIV-1 genotype B field isolate by nucleic acid amplification techniques

An international collaborative study to assess inter-laboratory variation in the sensitivity of gene amplification assays for the detection of HIV-1 RNA sequences was conducted using a panel of eight duplicate dilutions of an HIV-1 genotype B clinical isolate and negative control samples. Twenty-fiv...

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Bibliographic Details
Published in:Journal of virological methods 1999-03, Vol.78 (1), p.21-34
Main Authors: Bootman, J., Heath, A.B., Hughes, P., Holmes, H
Format: Article
Language:English
Subjects:
Adult
AIDS/HIV
Biological and medical sciences
DNA, Complementary
DNA, Viral - analysis
Fundamental and applied biological sciences. Psychology
HIV Core Protein p24 - analysis
HIV Infections - virology
HIV-1
HIV-1 - genetics
HIV-1 - isolation & purification
Humans
International Cooperation
Laboratories - standards
Male
Microbiology
Nucleic Acid Amplification Techniques
PCR
Polymerase Chain Reaction - standards
Reproducibility of Results
RNA, Viral - analysis
RNA, Viral - genetics
Sensitivity and Specificity
Standardisation
Techniques used in virology
Viral RNA
Virology
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Summary:An international collaborative study to assess inter-laboratory variation in the sensitivity of gene amplification assays for the detection of HIV-1 RNA sequences was conducted using a panel of eight duplicate dilutions of an HIV-1 genotype B clinical isolate and negative control samples. Twenty-five laboratories participated in the study and used a variety of in-house assays and commercial assay systems. With few exceptions, the assays were more sensitive than a p24 antigen assay. Overall, the PCR-based Amplicor Monitor assay was the most sensitive and gave the highest mean copy number for any one sample. Some of the in-house assays gave results comparable with the Monitor assay whilst the NASBA and bDNA assays appeared to be less sensitive. As a result of this study, an HIV-1 Working Reagent for the standardisation of nucleic acid amplification assays was developed and assessed in a subsequent study. Similar differences in sensitivity between the different assay systems was observed. The discrepancies in viral copy number obtained using the Working Reagent highlights the need for an International Standard against which all Working Reagents may be calibrated.
ISSN:0166-0934
1879-0984
DOI:10.1016/S0166-0934(98)00159-1