Near point-of-care, point-mutation test to detect drug resistance in HIV-1: a validation study in a Mexican cohort. (15th July 2020)

• Record Type: Journal Article
• Authors: Panpradist, Nuttada; Beck, Ingrid A.; Ruth, Parker S.; Ávila-Ríos, Santiago; García-Morales, Claudia; Soto-Nava, Maribel; Tapia-Trejo, Daniela; Matías-Florentino, Margarita; Paz-Juarez, Hector E.; del Arenal-Sanchez, Silvia; Reyes-Terán, Gustavo; Lutz, Barry R.; Frenkel, Lisa M.
• Is Part Of: AIDS. Volume 34:Number 9(2020)
• Publication Date: 2020-07-15
• Subjects: ART switching -- individualized medicine -- NNRT-based regimen -- OLA-Simple -- personalized medicine -- TLD switching; AIDS (Disease) -- Periodicals; Acquired Immunodeficiency Syndrome; AIDS (Disease); Periodicals; Periodicals; 616.9792005
• DOI: 10.1097/QAD.0000000000002524
• Languages: English
• ISSNs: 0269-9370

Abstract

Abstract : Objective: Pretreatment HIV-drug resistance (PDR, HIVDR) to non-nucleoside reverse transcriptase inhibitors (NNRTIs) is increasing globally. NNRTIs continue to be used as first-line antiretroviral therapy (ART) in some communities due to the cost of dolutegravir-based ART or dolutegravir-associated adverse events. A simplified version of the oligonucleotide ligation assay (OLA) – 'OLA-Simple' – is a low-cost, near point-of-care assay that provides ready-to-use lyophilized reagents and reports HIVDR mutations as colored lines on lateral flow strips. Our objective was to design and validate OLA-Simple for a Mexican cohort. Design: OLA-Simple probes to detect K65R, K103N/S, Y181C, M184V, and G190A were optimized for HIV Mexican sequences. Sixty clinical plasma specimens were analyzed by OLA-Simple by technicians blinded to Illumina-MiSeq sequences, and HIVDR results were compared. Methods: Plasma RNA was tested using OLA-Simple kits. OLA-Simple lateral flow strips were read by in-house software, and were classified as mutant or wild-type at each codon. The comparison of results by OLA-Simple and Miseq was used to generate receiver-operating characteristic curves. Results: OLA-Simple PCR amplified 59 of 60 specimens and successfully genotyped 287 of 295 codons, with eight of 295 (2.7%) indeterminate results. Compared to MiSeq, OLA-Simple gave five of 295 (1.7%) false-positive and four of 295 (1.4%) false-negative results. Excluding indeterminate results, OLA-Simple classified mutant with an accuracy of 97.4 and 98.8% when using thresholds at 10 and 25% mutant within an individual's HIV quasispecies, respectively. Conclusions: Compared to MiSeq, OLA-Simple detected HIVDR with high sensitivity and accuracy. OLA-Simple could expand access to affordable and rapid HIVDR testing to guide appropriate ART choices in populations using NNRTI-based ART. Abstract : Supplemental Digital Content is available in the text