In-house HIV-1 RNA real-time RT-PCR assays: principle, available tests and usefulness in developing countries.
The principle of currently available licensed HIV-1 RNA assays is based on real-time technologies that continuously monitor the fluorescence emitted by the amplification products. Besides these assays, in-house quantitative (q) real-time reverse transcription (RT)-PCR (RT-qPCR) tests have been developed and evaluated particularly in developing countries, for two main reasons. First, affordable and generalized access to HIV-1 RNA viral load is urgently needed in the context of expected universal access to prevention and antiretroviral treatment programs in these settings. Second, since many non-B subtypes, circulating recombinant forms and unique recombinant forms circulate in these areas, in-house HIV-1 RNA RT-qPCR assays are ideal academic tools to thoroughly evaluate the impact of HIV-1 genetic diversity on the accuracy of HIV-1 RNA quantification, as compared with licensed techniques. To date, at least 15 distinct in-house assays have been designed. They differ by their chemistry and the HIV-1 target sequence (located in gag, Pol-IN or LTR gene). Analytical performances of the tests that have been extensively evaluated appear at least as good as (or even better than) those of approved assays, with regard to HIV-1 strain diversity. Their clinical usefulness has been clearly demonstrated for early diagnosis of pediatric HIV-1 infection and monitoring of highly active antiretroviral therapy efficacy. The LTR-based HIV-1 RNA RT-qPCR assay has been evaluated by several groups under the auspices of the Agence Nationale de Recherches sur le SIDA et les hépatites virales B et C. It exists now as a complete standardized commercial test.
Rouet
François
F
Laboratoire de Virologie, Centre Muraz, BP390 Bobo-Dioulasso 01, Burkina Faso. franrouet@yahoo.fr
Ménan
Hervé
H
Viljoen
Johannes
J
Ngo-Giang-Huong
Nicole
N
Mandaliya
Kishor
K
Valéa
Diane
D
Lien
Truong Xuan
TX
Danaviah
Sivapragashini
S
Rousset
Dominique
D
Ganon
Amandine
A
Nerrienet
Eric
E
D017931:DNA Primers / Q000235:genetics; D003906:Developing Countries; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013997:Time Factors
D017931:DNA Primers; D012367:RNA, Viral