Analytical Performances of Human Immunodeficiency Virus Type 1 RNA-Based Amplix® Real-Time PCR Platform for HIV-1 RNA Quantification.

Laurent Bélec,Mathieu Matta,Mohammad-Ali Jenabian,Christian Diamant Mossoro-Kpinde,Laurent Andreoletti,Jean De Dieu Longo,Serge Tonen Wolyec,Leman Robin,Gérard Grésenguet,Ralph-Sydney Mboumba Bouassa

doi:10.1155/2016/7954810

Abstract

Objectives. We evaluated the performances of Amplix real-time PCR platform developed by Biosynex (Strasbourg, France), combining automated station extraction (Amplix station 16 Dx) and real-time PCR (Amplix NG), for quantifying plasma HIV-1 RNA by lyophilized HIV-1 RNA-based Amplix reagents targeting gag and LTR, using samples from HIV-1-infected adults from Central African Republic. Results. Amplix real-time PCR assay showed low limit of detection (28 copies/mL), across wide dynamic range (1.4–10 log copies/mL), 100% sensitivity and 99% specificity, high reproducibility, and accuracy with mean bias < 5%. The assay showed excellent correlations and concordance of 95.3% with the reference HIV-1 RNA load assay (Roche), with mean absolute bias of +0.097 log copies/mL by Bland-Altman analysis. The assay was able to detect and quantify the most prevalent HIV-1 subtype strains and the majority of non-B subtypes, CRFs of HIV-1 group M, and HIV-1 groups N and O circulating in Central Africa. The Amplix assay showed 100% sensitivity and 99.6% specificity to diagnose virological failure in clinical samples from antiretroviral drug-experienced patients. Conclusions. The HIV-1 RNA-based Amplix real-time PCR platform constitutes sensitive and reliable system for clinical monitoring of HIV-1 RNA load in HIV-1-infected children and adults, particularly adapted to intermediate laboratory facilities in sub-Saharan Africa.

Highlights

The increased availability of antiretroviral treatment (ART) for HIV infection has resulted in major reductions in morbidity and mortality in high HIV burden with poor resources settings
We evaluated the HIV-1 RNA-based Amplix real-time PCR platform according to the Clinical and Laboratory Standards Institute (CLSI) guidelines for clinical use [10, 11] and guidelines recommended by the pSMILE project (Patient Safety Monitoring in International Laboratories), in Bangui, the capital of the Central African Republic, a country of broad genetic diversity and high prevalence of non-B subtypes of HIV-1 group M
Plasma HIV-1 RNA load is a routine investigation for monitoring of HIV-1-infected individuals

Summary

Introduction

The increased availability of antiretroviral treatment (ART) for HIV infection has resulted in major reductions in morbidity and mortality in high HIV burden with poor resources settings. The 2013revised WHO guidelines for scaling up ART in adults and children living in resource-limited settings emphasized the need of laboratory monitoring, first based on immunological assessment of CD4 T-cell count, mainly to start ART and monitor patients on ART, and secondly based on the plasma HIV-1 RNA load in order to monitor treatment efficacy and early therapeutic failure as well as to monitor therapeutic switches [2,3,4,5]. A comprehensive revision of the consolidated WHO guidelines on the use of ART has been undertaken in 2015 based on new scientific evidence and lessons from antiretroviral programs implementation [1]. The key recommendation made in 2015 is the fact that ART should be initiated in everyone living with HIV at any CD4 T-cell count as this may result in a better clinical outcome [1]

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Conclusion