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Therapeutic Methods and Therapies TCIM
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J Virol Methods ; 243: 151-157, 2017 05.
Article in English | MEDLINE | ID: mdl-27894862

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.


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