title: In-house HIV-1 RNA real-time RT-PCR assays: principle, available tests and usefulness in developing countries.
abstract: 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.
authors.affiliation: Laboratoire de Virologie, Centre Muraz, BP390 Bobo-Dioulasso 01, Burkina Faso. franrouet@yahoo.fr
authors.forename: François
authors.initials: F
authors.lastname: Rouet
authors.forename: Hervé
authors.initials: H
authors.lastname: Ménan
authors.forename: Johannes
authors.initials: J
authors.lastname: Viljoen
authors.forename: Nicole
authors.initials: N
authors.lastname: Ngo-Giang-Huong
authors.forename: Kishor
authors.initials: K
authors.lastname: Mandaliya
authors.forename: Diane
authors.initials: D
authors.lastname: Valéa
authors.forename: Truong Xuan
authors.initials: TX
authors.lastname: Lien
authors.forename: Sivapragashini
authors.initials: S
authors.lastname: Danaviah
authors.forename: Dominique
authors.initials: D
authors.lastname: Rousset
authors.forename: Amandine
authors.initials: A
authors.lastname: Ganon
authors.forename: Eric
authors.initials: E
authors.lastname: Nerrienet
mesh_terms: 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
chemical_list: D017931:DNA Primers; D012367:RNA, Viral