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Journal of Clinical Microbiology, June 2005, p. 2709-2717, Vol. 43, No. 6
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.6.2709-2717.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Transfer and Evaluation of an Automated, Low-Cost Real-Time Reverse Transcription-PCR Test for Diagnosis and Monitoring of Human Immunodeficiency Virus Type 1 Infection in a West African Resource-Limited Setting

Francois Rouet,1* Didier K. Ekouevi,2 Marie-Laure Chaix,3 Marianne Burgard,3 Andre Inwoley,1 Thomas D'Aquin Tony,4 Christine Danel,5 Xavier Anglaret,5,6 Valeriane Leroy,2,6 Philippe Msellati,7 Francois Dabis,2,6 and Christine Rouzioux3

Centre de Diagnostic et de Recherches sur le SIDA, Abidjan, Côte d'Ivoire,1 ANRS 1201/1202 Ditrame Plus Program, Abidjan, Côte d'Ivoire,2 Laboratoire de Virologie, EA 3620 Université René Descartes, C. H. U. Necker, Enfants Malades, Paris, France,3 ANRS 1220 Primo-ci Cohort, Abidjan, Côte d'Ivoire,4 ANRS 1269 Trivacan Trial, Abidjan, Côte d'Ivoire,5 INSERM U593, ISPED, Université Victor Segalen, Bordeaux, France,6 ANRS 1244/1278 Pediatric Program, Abidjan, Côte d'Ivoire7

Received 4 October 2004/ Returned for modification 31 December 2004/ Accepted 25 January 2005

There is an urgent need for low-cost human immunodeficiency virus type 1 (HIV-1) viral load (VL) monitoring technologies in resource-limited settings. An automated TaqMan real-time reverse transcription-PCR (RT-PCR) assay was transferred to the laboratory of the Centre de Diagnostic et de Recherches sur le SIDA, Abidjan, Côte d'Ivoire, and assessed for HIV-1 RNA VL testing in 806 plasma samples collected within four ANRS research programs. The detection threshold and reproducibility of the assay were first determined. The quantitative results obtained with this assay were compared with two commercial HIV-1 RNA kits (the Versant version 3.0 and Monitor version 1.5 assays) in specimens harboring mainly the circulating recombinant form 02 strain (CRF02). The clinical evaluation of this test was done in different situations including the early diagnosis of pediatric infection and the monitoring of antiretroviral-treated patients. The quantification limit of our method was 300 copies/ml. The HIV-1 RNA values obtained by real-time PCR assay were highly correlated with those obtained by the Versant kit (r = 0.901; P < 0.001) and the Monitor test (r = 0.856; P < 0.001) and homogeneously distributed according to HIV-1 genotypes. For the early diagnosis of pediatric HIV-1 infection, the sensitivity and specificity of the real-time PCR assay were both 100% (95% confidence intervals of 93.7 to 100.0 and 98.3 to 100.0, respectively), compared to the Versant results. Following initiation of antiretroviral treatment, the kinetics of HIV-1 RNA levels were very comparable, with a similar proportion of adults and children below the detection limit during follow-up with our technique and the Versant assay. The TaqMan real-time PCR ($12 per test) is now routinely used to monitor HIV-1 infection in our laboratory. This technology should be further evaluated in limited-resource countries where strains other than CRF02 are prevalent.

* Corresponding author. Present address: 24, rue du Centre, 85410 Thouarsais-Bouildroux, France. Phone and fax: 33 02 51 51 3899. E-mail: rouet{at}aviso.ci.

Journal of Clinical Microbiology, June 2005, p. 2709-2717, Vol. 43, No. 6
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.6.2709-2717.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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