A single-loop recombinant pseudotyped-virus-based assay to detect HIV-1 phenotypic resistance

Area of Science:

• Virology
• Molecular Biology
• Public Health

Background:

• HIV/AIDS remains a global health challenge.
• Highly active antiretroviral therapy (HAART) is the standard treatment for HIV/AIDS.
• Emerging drug resistance in HIV-1 strains compromises long-term ART effectiveness.

Purpose of the Study:

• To develop a single-loop recombinant pseudotyped-virus-based assay for detecting phenotypic resistance in clinical HIV-1 strains.
• To provide a tool for monitoring HIV-1 drug resistance and evaluating antiretroviral agent sensitivity.

Main Methods:

• Extracted HIV-1 RNA from patient plasma and amplified key viral genes (protease, reverse transcriptase, integrase).
• Constructed resistance test vectors (RTVs) using a retroviral vector and luciferase reporter.
• Produced pseudotyped virus stocks and monitored viral replication via luciferase activity in infected cells.

Main Results:

• Successfully amplified target fragments from 97.1% of clinical samples and constructed RTVs from 94.3%.
• Demonstrated a clear dose-dependent relationship between virus production, luciferase activity, and drug concentration.
• Phenotypic resistance profiles closely matched observed genotypes in 29 clinical samples.

Conclusions:

• A stable, single-loop recombinant pseudotyped-virus assay for phenotypic HIV-1 resistance testing was successfully developed.
• This assay is applicable for evaluating clinical HIV-1 strains' resistance to antiretroviral drugs.
• The assay aids in monitoring treatment effectiveness and guiding therapeutic decisions for HIV/AIDS patients.