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Reproducibility of a high-throughput HIV-1 genotypic resistance assay over time (2001–2007)
Journal of the International AIDS Society volume 11, Article number: P205 (2008)
Designed for innovative HIV-1 diagnostics and resistance analysis, a high-throughput laboratory-developed (Protease-Reverse-Transcriptase) genotyping assay was implemented at Virco (Belgium). Following regulatory guidelines, assay variability was monitored over time by means of a quarterly internal quality control (QC) panel.
The panel comprises seven well-characterized recombinant viruses that were derived from clinical isolates. Virus stocks were grown and later re-cultured as needed and aliquoted for use at each quarterly testing between Q4 in 2001 through Q1 in 2007. The nucleotide similarity among the original virus stocks and subsequent re-cultured viruses, was determined using pairwise BLAST analysis (BioEdit Sequence Alignment Editor).
Summary of results
A total of 603 and 867 pairwise BLAST comparisons were made, yielding an average within and between original and re-cultured virus stocks similarity of 99.60% ± 0.27 and 99.52% ± 0.33, respectively.
Since both BLAST values did not differ, it can be concluded that re-culturing of viruses did not lead to a significant decrease in similarity. Furthermore, despite the presence of high levels of nucleotide sequence mixtures of the original clinical isolates, the high reproducibility of the genotyping assay (more than 99% sequence identity) was demonstrated. A combination of optimal assay design, rigorous personnel training, the testing of the internal QC panel at regular intervals and participation in a CLIA-approved 6-monthly external proficiency testing program (Accutest) ensures that quality standards are consistently met for the genotyping assay performed at Virco.
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Deloof, A., Muyldermans, G., Pattery, T. et al. Reproducibility of a high-throughput HIV-1 genotypic resistance assay over time (2001–2007). JIAS 11, P205 (2008). https://doi.org/10.1186/1758-2652-11-S1-P205
- Proficiency Testing
- Genotyping Assay
- Virus Stock
- Internal Quality Control
- BioEdit Sequence Alignment Editor