Development and genomic characterization of a diverse HIV-1 variant reference panel for nucleic acid-based testing

Area of Science:

• Virology
• Molecular Biology
• Public Health

Background:

• Global Human Immunodeficiency Virus (HIV) genetic diversity complicates accurate detection and diagnosis via nucleic acid testing (NAT).
• Well-characterized reference panels are crucial for assessing the analytical performance of HIV diagnostic tests.

Purpose of the Study:

• To develop a comprehensive reference panel for HIV NAT that accurately represents the genetic diversity of circulating HIV strains.
• To create a standardized tool for validating HIV detection methods.

Main Methods:

• HIV-1 strains were cultured from clinical specimens in Cameroon.
• Near full-length genomes were sequenced using metagenomic next-generation sequencing and Sanger sequencing.
• A diverse subset of 18 HIV-1 strains was selected to form the reference panel, with viral loads quantified at low, medium, and high levels.

Main Results:

• Near full-length HIV-1 genomes (average 9589 bp) were obtained for 37 out of 59 strains.
• The panel includes 18 diverse HIV-1 strains: 4 subtypes, 4 circulating recombinant forms, and 8 unique recombinant forms.
• Panel members were prepared at three viral load concentrations (low, medium, high), resulting in 51 total panel members.

Conclusions:

• The developed HIV reference panel effectively represents diverse circulating strains.
• This panel is a valuable resource for evaluating the performance of HIV NAT assays.
• The panel is available to developers and manufacturers of HIV diagnostic tests upon request.