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.

A signal amplification system on a lateral flow immunoassay detecting for hepatitis e‐antigen in human blood samples

“…Ninety‐nine plasma samples with both HBsAg and HBeAg positive and 291 plasma samples with negative HBsAg were obtained from blood donors in Guangzhou Blood Center, which were routinely screened by ELISA and Utrio Plus nucleic acid testing (individual‐donation NAT, Grifols Diagnostic Solutions, San Diego, CA; limit of detection HBV DNA 3.4 IU/mL), respectively . The non‐HBV infected sera or plasmas of hepatitis C virus (HCV) infected patients, human immunodeficiency virus (HIV) infected patients or rabies vaccine immunized patients from the laboratory were used as controls for testing of assay specificity . This study was approved by the Medical Ethics Committees of Southern Medical University and Guangzhou Blood Center and followed the ethical guidelines of the 1975 Declaration of Helsinki.…”

Evaluation of the ABL NGS assay for HIV-1 drug resistance testing

“…HIV's resistance to various drug molecules can be assessed by phenotypic or genotypic testing. Phenotypic testing is reserved for highly specialized HIV research laboratories or require expertise, whereas genotypic tests are commonly used in laboratories monitoring infected patients [ 4 , 5 ]. In genotypic testing, the target genetic material is amplified in PCRs and the region of interest is then sequenced for mutations known to confer resistance to antiretrovirals.…”