TY - JOUR
T1 - Development and evaluation of an assay for HIV-1 protease and reverse transcriptase drug resistance genotyping of all major group-M subtypes
AU - Aitken, Susan C.
AU - Kliphuis, Aletta
AU - Wallis, Carole L.
AU - Chu, Mei Ling
AU - Fillekes, Quirine
AU - Barth, Roos
AU - Stevens, Wendy
AU - Rinke de Wit, Tobias F.
AU - Schuurman, Rob
PY - 2012
Y1 - 2012
N2 - Background: High cost and varying sensitivity for non-B HIV-1 subtypes limits application of current commercial kits for HIV-1 drug resistance genotyping of all major HIV-1 group-M subtypes. Objectives: Our research aimed to develop and validate an assay specific for all major HIV-1 group-M subtypes for use as an alternative to commercial assays for HIV-1 protease (PR) and reverse transcriptase (RT) drug resistance genotyping. Study design: A nested RT-PCR encompassing the entire PR and RT up to amino acid 321 of HIV-1 was designed to detect HIV-1 group-M subtypes. Primers compatible with group-M subtypes were defined and analytical sensitivity of the assay evaluated using a panel of reference viruses for subtypes A-H and CRF01_AE. The assay was subsequently evaluated on 246 plasma samples from HIV-1 infected individuals harboring various group-M subtypes and viral loads (VLs). Results: All major group-M HIV-1 subtypes were detected with an overall analytical sensitivity of 1.00E+03 RNA copies/ml. Application of the genotyping assay on 246 primarily African clinical samples comprising subtypes A (n = 52; 21.7%), B (n = 12; 5.0%), C (n = 127; 52.9%), D (n = 25; 10.4%), CRF01_AE (n = 10; 4.2%), and CRF02_AG (n = 10; 4.2%), and unassigned variants (n = 10; 4.2%), VL range 4.32E+02-8.63E+06 (median 2.66E+04) RNA copies/ml, was similar to 98% successful. Conclusions: A group-M subtype-independent genotyping assay for detection of HIV-1 drug resistance was developed. The described assay can serve as an alternative to commercial assays for HIV-1 drug resistance genotyping in routine diagnostics, and for surveillance and monitoring of drug resistance in resource-limited settings (RLS). (C) 2012 Elsevier B. V. All rights reserved
AB - Background: High cost and varying sensitivity for non-B HIV-1 subtypes limits application of current commercial kits for HIV-1 drug resistance genotyping of all major HIV-1 group-M subtypes. Objectives: Our research aimed to develop and validate an assay specific for all major HIV-1 group-M subtypes for use as an alternative to commercial assays for HIV-1 protease (PR) and reverse transcriptase (RT) drug resistance genotyping. Study design: A nested RT-PCR encompassing the entire PR and RT up to amino acid 321 of HIV-1 was designed to detect HIV-1 group-M subtypes. Primers compatible with group-M subtypes were defined and analytical sensitivity of the assay evaluated using a panel of reference viruses for subtypes A-H and CRF01_AE. The assay was subsequently evaluated on 246 plasma samples from HIV-1 infected individuals harboring various group-M subtypes and viral loads (VLs). Results: All major group-M HIV-1 subtypes were detected with an overall analytical sensitivity of 1.00E+03 RNA copies/ml. Application of the genotyping assay on 246 primarily African clinical samples comprising subtypes A (n = 52; 21.7%), B (n = 12; 5.0%), C (n = 127; 52.9%), D (n = 25; 10.4%), CRF01_AE (n = 10; 4.2%), and CRF02_AG (n = 10; 4.2%), and unassigned variants (n = 10; 4.2%), VL range 4.32E+02-8.63E+06 (median 2.66E+04) RNA copies/ml, was similar to 98% successful. Conclusions: A group-M subtype-independent genotyping assay for detection of HIV-1 drug resistance was developed. The described assay can serve as an alternative to commercial assays for HIV-1 drug resistance genotyping in routine diagnostics, and for surveillance and monitoring of drug resistance in resource-limited settings (RLS). (C) 2012 Elsevier B. V. All rights reserved
U2 - https://doi.org/10.1016/j.jcv.2012.01.010
DO - https://doi.org/10.1016/j.jcv.2012.01.010
M3 - Article
C2 - 22306272
SN - 1386-6532
VL - 54
SP - 21
EP - 25
JO - Journal of clinical virology
JF - Journal of clinical virology
IS - 1
ER -