Assessing the Variability of Cell-Associated HIV DNA Quantification through a Multicenter Collaborative Study.
Yann Le DuffKathleen GärtnerEloise J BusbyAnnalisa DalziniSivapragashini DanaviahJosé Luis Jiménez FuentesCarlo GiaquintoJim F HuggettMatthew HurleyAnne-Geneviève MarcellinMaría Ángeles Muñoz-FernándezDenise M O'SullivanDeborah PersaudLaura PowellPeter RigsbyPaolo RossiAnita de RossiLilly SiemsTheresa SmitSarah A WattersNeil AlmondEleni Nastoulinull nullPublished in: Microbiology spectrum (2022)
Reliable and accurate quantification of cell-associated HIV DNA (CA HIV DNA) is critical for early infant diagnosis, clinical management of patients under therapy, and to inform new therapeutics efficacy. The present study assessed the variability of CA HIV DNA quantification obtained from various assays and the value of using reference materials to help harmonize the measurements. Using a common set of reagents, our multicenter collaborative study highlights significant variability of CA HIV DNA quantification and lower limit of quantification across assays. The quantification of CA HIV DNA from a panel of infected PBMCs can be harmonized through cross-subtype normalization but assay calibration with the commonly used 8E5 cell line failed to reduce quantification variability between assays, demonstrating the requirement to thoroughly evaluate reference material candidates to help improve the comparability of CA HIV DNA diagnostic assay performance. IMPORTANCE Despite a global effort, HIV remains a major public health burden with an estimated 1.5 million new infections occurring in 2020. HIV DNA is an important viral marker, and its monitoring plays a critical role in the fight against HIV: supporting diagnosis in infants and underpinning clinical management of patients under therapy. Our study demonstrates that HIV DNA measurement of the same samples can vary significantly from one laboratory to another, due to heterogeneity in the assay, protocol, and reagents used. We show that when carefully selected, reference materials can reduce measurement variability and harmonize HIV DNA quantification across laboratories, which will help contribute to improved diagnosis and clinical management of patients living with HIV.
Keyphrases
- antiretroviral therapy
- hiv positive
- hiv testing
- hiv infected
- human immunodeficiency virus
- hepatitis c virus
- hiv aids
- men who have sex with men
- circulating tumor
- single molecule
- cell free
- public health
- high throughput
- south africa
- randomized controlled trial
- cell therapy
- mass spectrometry
- high resolution
- circulating tumor cells
- risk factors
- quality improvement
- protein kinase
- double blind