Dry Panels Supporting External Quality Assessment Programs for Next Generation Sequencing-Based HIV Drug Resistance...

Area of Science:

• Genomics
• Virology
• Bioinformatics

Background:

• External quality assessment (EQA) is vital for validating next-generation sequencing (NGS)-based HIV drug resistance testing (DRT).
• While generating NGS data for HIV-DRT is increasingly accessible, bioinformatic analysis remains a significant challenge for many diagnostic labs.
• Existing software tools for analyzing NGS data show discrepancies, especially for low-frequency variations.

Purpose of the Study:

• To highlight the critical need for validating bioinformatics tools used in NGS HIV-DRT.
• To emphasize the importance of well-characterized reference datasets ('dry panels') for accurate software evaluation.
• To present criteria for designing effective dry panels to support EQA programs.

Main Methods:

• Discussion and consensus building during the Second International Winnipeg Symposium on EQA strategies for NGS HIV-DR assays.
• Identification of challenges in bioinformatic data analysis for HIV-DRT.
• Proposal of criteria for the design of 'dry panels'.

Main Results:

• Bioinformatic analysis of NGS data for HIV-DRT presents a bottleneck due to software discrepancies, particularly with low-frequency variations.
• The development of standardized 'dry panels' with known ground truth is essential for validating bioinformatics tools.
• Criteria for designing these dry panels were discussed and formulated.

Conclusions:

• Robust validation of bioinformatics tools is critical for reliable NGS-based HIV-DRT.
• The strategic design of dry panels is imperative for advancing EQA in this field.
• These efforts will improve the accuracy and consistency of HIV drug resistance testing globally.