A comparative evaluation of the performance of the Viroseq HIV-1 genotyping system v2.0 against an in-house assay using specimens from the national HIV drug resistance monitoring survey, in Zimbabwe.

摘要: This study was carried out to assess the performance of Viroseq HIV-1 genotyping system v2.0 (Celera, Alameda, California, USA) on Zimbabwean samples at National Microbiology Reference Laboratory (NMRL) in Harare and compare results obtained with those obtained a World Health Organization accredited genotyping laboratory in Entebbe, Uganda, which genotyped same using an in-house method. Study samples: Thirty eight (38) plasma were used this study. They selected from stored NMRL, had been collected from patients eligible for commencement antiretroviral therapy just prior taking drugs sentinel sites for National HIV Drug Resistance Monitoring Survey Zimbabwe. Results: Viral RNA extraction RT-PCR reactions successful first attempt all the 38 samples. The success rates cycle sequencing 7 proprietary Viroseq primer mixes samples, were: Primer A (81.6%), primer B (86.8), primers C G (84.2%), D (10.5%), F (73.7%) Primer H (94.7%). seven (97.4%) successfully genotyped. However, 14 (37.8%) these pol region consensus sequences unidirectional. Thirty two 37 (86.5%) clustered pairs with their corresponding sample generated Uganda bootstrap support values ≥70 (median value, 97.5: range 31-99) neighbour joining tree phylogenetic analysis. All found be subtype (n=37). Nineteen (48.6%) least one drug resistance mutation detected by either or Ugandan assay. detected mutations as reported by systems fully concordant 11/19 (57.9%), partially concordant in 2/19 (10.5%) discordant 6/19 (31.6%) specimens resistance mutations. Overall, all reports without any mutations 78.4% concordant, 16.2% 5.4% discordant. Conclusions: system can reliably genotype by adequately trained operators NMRL. Sequencing is not suitable sequencing Zimbabwean isolates due its high reaction failure rate.