title: Use of amplification refractory mutation system PCR assay as a simple and effective tool to detect HIV-1 drug resistance mutations.
abstract: Access to genotyping assays to determine successful antiretroviral treatment (ART) is limited in resource-constrained settings by high cost, suggesting the need for a cost-effective and simplified method to identify HIV-1 drug resistance (HIVDR) mutations. In this study, an amplification refractory mutation system (ARMS)-PCR assay was developed and used to investigate the most frequent HIVDR mutations affecting first-line ART in settings where WHO ART guidelines are applied. Seventy-five HIV-positive (HIV(+)) samples from Cameroon were used to assess the performance of this assay. Sequencing of HIV-1 reverse transcriptase was simultaneously performed for comparison, and discordant samples were tested with a Trugene HIV-1 genotyping kit. The ARMS-PCR assay was able to detect M184V, T215Y/F, K103N, and Y181C mutations with sensitivities of 96.8%, 85.7%, 91.3%, and 70%, respectively, and specificities of 90.6%, 95%, 100%, 96.9%, respectively, compared with data on sequencing. The results indicated the highest positive predictive value for K103N (100%) and the highest negative predictive value for M184V (97.5%). ARMS-PCR's limits of detection for mutations M184V, T215Y/F, K103N, and Y181C were <75 copies/ml, 143 copies/ml, 143 copies/ml, and 836 copies/ml, respectively. ARMS-PCR efficiently identified mutations in individuals harboring different HIV-1 clades (CRF02_AG and non-CRF02_AG). In addition, this approach was more cost-effective than other genotyping assays. The high throughput, the cost-effectiveness, and the simplicity of the ARMS-PCR assay make it a suitable tool to monitor HIVDR patterns in resource-constrained settings with broad HIV-1 genetic diversity.
authors: Department of Pathology, New York University School of Medicine, New York, New York, USA Department of Immunology and Applied Biotechnology, University of Rome Tor Vergata, Rome, Italy Chantal Biya International Reference Center, Sequencing Unit, Yaounde, Cameroon.
authors: Aubin J
authors: AJ
authors: Nanfack
authors: Faculty of Sciences, University of Dschang, Dschang, Cameroon Serology Unit, Medical Diagnostic Center, Yaounde, Cameroon.
authors: Lucy
authors: L
authors: Agyingi
authors: Serology Unit, Medical Diagnostic Center, Yaounde, Cameroon Internal Medicine Unit, Edéa Regional Hospital, Edéa, Cameroon.
authors: Jean Jacques N
authors: JJ
authors: Noubiap
authors: Serology Unit, Medical Diagnostic Center, Yaounde, Cameroon.
authors: Johnson N
authors: JN
authors: Ngai
authors: Department of Immunology and Applied Biotechnology, University of Rome Tor Vergata, Rome, Italy UNESCO Chair of Interdisciplinary Biotechnology, University of Rome Tor Vergata, Rome, Italy.
authors: Vittorio
authors: V
authors: Colizzi
authors: Department of Pathology, New York University School of Medicine, New York, New York, USA Veterans Affairs New York Harbor Healthcare Systems, New York, New York, USA phillipe.nyambi@nyumc.org.
authors: Phillipe N
authors: PN
authors: Nyambi
mesh_terms: D000328:Adult; D002163:Cameroon; D003362:Cost-Benefit Analysis; D024882:Drug Resistance, Viral; D005260:Female; D060005:Genotyping Techniques / Q000191:economics / Q000379:methods*; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D008297:Male; D008826:Microbial Sensitivity Tests / Q000191:economics / Q000379:methods*; D008875:Middle Aged; D020125:Mutation, Missense; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity