HIV drug resistance testing among patients failing second line antiretroviral therapy. Comparison of in-house and commercial sequencing

J Virol Methods. 2017 May:243:151-157. doi: 10.1016/j.jviromet.2016.11.010. Epub 2016 Nov 25.

Abstract

Introduction: HIV genotyping is often unavailable in low and middle-income countries due to infrastructure requirements and cost. We compared genotype resistance testing in patients with virologic failure, by amplification of HIV pol gene, followed by "in-house" sequencing and commercial sequencing.

Methods: Remnant plasma samples from adults and children failing second-line ART were amplified and sequenced using in-house and commercial di-deoxysequencing, and analyzed in Harare, Zimbabwe and at Stanford, U.S.A, respectively. HIV drug resistance mutations were determined using the Stanford HIV drug resistance database.

Results: Twenty-six of 28 samples were amplified and 25 were successfully genotyped. Comparison of average percent nucleotide and amino acid identities between 23 pairs sequenced in both laboratories were 99.51 (±0.56) and 99.11 (±0.95), respectively. All pairs clustered together in phylogenetic analysis. Sequencing analysis identified 6/23 pairs with mutation discordances resulting in differences in phenotype, but these did not impact future regimens.

Conclusions: The results demonstrate our ability to produce good quality drug resistance data in-house. Despite discordant mutations in some sequence pairs, the phenotypic predictions were not clinically significant.

Keywords: Genotyping; HIV drug resistance; SATuRN; Treatment failure; Zimbabwe.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Anti-Retroviral Agents / pharmacology
  • Anti-Retroviral Agents / therapeutic use*
  • Antiretroviral Therapy, Highly Active
  • Drug Resistance, Viral
  • Genotyping Techniques / methods*
  • HIV / drug effects*
  • HIV / genetics
  • HIV / isolation & purification
  • HIV Infections / drug therapy*
  • HIV Infections / virology*
  • Humans
  • Microbial Sensitivity Tests / methods
  • Sequence Analysis, DNA / methods*
  • Treatment Failure
  • United States
  • Zimbabwe

Substances

  • Anti-Retroviral Agents