title: A simple and cost-saving phenotypic drug susceptibility testing of HIV-1.
abstract: It is essential to monitor the occurrence of drug-resistant strains and to provide guidance for clinically adapted antiviral treatment of HIV/AIDS. In this study, an individual patient's HIV-1 pol gene encoding the full length of protease and part of the reverse transcriptase was packaged into a modified lentivirus carrying dual-reporters ZsGreen and luciferase. The optimal coefficient of correlation between drug concentration and luciferase activity was optimized. A clear-cut dose-dependent relationship between lentivirus production and luciferase activity was found in the phenotypic testing system. Fold changes (FC) to a wild-type control HIV-1 strain ratios were determined reflecting the phenotypic susceptibility of treatment-exposed patient's HIV-1 strains to 12 HIV-1 inhibitors including 6 nucleoside reverse-transcriptase inhibitors (NRTIs), 4 non-nucleoside reverse transcriptase inhibitors (NNRTIs) and 2 protease inhibitors (PIs). Phenotypic susceptibility calls from 8 HIV-1 infected patients were consistent with 80-90% genotypic evaluations, while phenotypic assessments rectified 10-20% genotypic resistance calls. By a half of replacement with ZsGreen reporter, the consumption of high cost Bright-Glo Luciferase Assay is reduced, making this assay cheaper when a large number of HIV-1 infected individuals are tested. The study provides a useful tool for interpreting meaningful genotypic mutations and guiding tailored antiviral treatment of HIV/AIDS in clinical practice.
authors.affiliation: Department of Transfusion Medicine, Southern Medical University, Guangzhou, China.
authors.forename: Yunceng
authors.initials: Y
authors.lastname: Weng
authors.affiliation: Department of Transfusion Medicine, Southern Medical University, Guangzhou, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Zhang
authors.affiliation: First Clinical Medicine School, Southern Medical University, Guangzhou, China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Huang
authors.affiliation: Shenzhen Center of Disease Prevention and Control, Shenzhen, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Department of Transfusion Medicine, Southern Medical University, Guangzhou, China.
authors.forename: Peifang
authors.initials: P
authors.lastname: Luo
authors.affiliation: Shenzhen Bioeasy Company, Shenzhen, China.
authors.forename: Siyuan
authors.initials: S
authors.lastname: Bi
authors.affiliation: Shenzhen Center of Disease Prevention and Control, Shenzhen, China.
authors.forename: Zhengrong
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Shenzhen Bioeasy Company, Shenzhen, China.
authors.forename: Hai
authors.initials: H
authors.lastname: Zhu
authors.affiliation: Department of Transfusion Medicine, Southern Medical University, Guangzhou, China.|Department of Haematology, University of Cambridge, UK.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Allain
authors.affiliation: Department of Transfusion Medicine, Southern Medical University, Guangzhou, China.|School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou, China.
authors.forename: Chengyao
authors.initials: C
authors.lastname: Li
mesh_terms: D019380:Anti-HIV Agents / Q000032:analysis* / Q000494:pharmacology*; D002452:Cell Count; D004353:Drug Evaluation, Preclinical / Q000191:economics* / Q000379:methods*; D024882:Drug Resistance, Viral / Q000187:drug effects*; D005838:Genotype; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D011995:Recombination, Genetic / Q000235:genetics; D015203:Reproducibility of Results; D013997:Time Factors
chemical_list: D019380:Anti-HIV Agents