HIV-1 phenotypic reverse transcriptase inhibitor drug resistance test interpretation is not dependent on the subtype of the virus backbone

• Published in: PloS one 2012-04, Vol.7 (4), p.e34708-e34708
• Main Authors: Bronze, Michelle, Steegen, Kim, Wallis, Carole L, De Wolf, Hans, Papathanasopoulos, Maria A, Van Houtte, Margriet, Stevens, Wendy S, de Wit, Tobias Rinke, Stuyver, Lieven J
• Format: Article
• Language: English
• Subjects: Acquired immune deficiency syndrome; AIDS; Analysis; Anti-HIV Agents - therapeutic use; Antiretroviral agents; Antiretroviral drugs; Backbone; Biology; Confidence intervals; Disease resistance; DNA polymerases; Drug resistance; Drug Resistance, Multiple, Viral; Ethics; Highly active antiretroviral therapy; HIV; HIV Infections - blood; HIV Infections - drug therapy; HIV Reverse Transcriptase - antagonists & inhibitors; HIV-1 - drug effects; HIV-1 - genetics; Human immunodeficiency virus; Humans; Medical screening; Medicine; Mutation; Phenotyping; Protease inhibitors; Reverse Transcriptase Inhibitors - therapeutic use; RNA-directed DNA polymerase; Variability; Viruses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0034708

To date, the majority of HIV-1 phenotypic resistance testing has been performed with subtype B virus backbones (e.g. HXB2). However, the relevance of using this backbone to determine resistance in non-subtype B HIV-1 viruses still needs to be assessed. From 114 HIV-1 subtype C clinical samples (36 ARV-naïve, 78 ARV-exposed), pol amplicons were produced and analyzed for phenotypic resistance using both a subtype B- and C-backbone in which the pol fragment was deleted. Phenotypic resistance was assessed in resulting recombinant virus stocks (RVS) for a series of antiretroviral drugs (ARV's) and expressed as fold change (FC), yielding 1660 FC comparisons. These Antivirogram® derived FC values were categorized as having resistant or sensitive susceptibility based on biological cut-off values (BCOs). The concordance between resistance calls obtained for the same clinical sample but derived from two different backbones (i.e. B and C) accounted for 86.1% (1429/1660) of the FC comparisons. However, when taking the assay variability into account, 95.8% (1590/1660) of the phenotypic data could be considered as being concordant with respect to their resistance call. No difference in the capacity to detect resistance associated with M184V, K103N and V106M mutations was noted between the two backbones. The following was concluded: (i) A high level of concordance was shown between the two backbone phenotypic resistance profiles; (ii) Assay variability is largely responsible for discordant results (i.e. for FC values close to BCO); (iii) Confidence intervals should be given around the BCO's, when assessing resistance in HIV-1 subtype C; (iv) No systematic resistance under- or overcalling of subtype C amplicons in the B-backbone was observed; (v) Virus backbone subtype sequence variability outside the pol region does not contribute to phenotypic FC values. In conclusion the HXB2 virus backbone remains an acceptable vector for phenotyping HIV-1 subtype C pol amplicons.