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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
medicine
Real-time PCR quantification of genital shedding of herpes simplex virus (HSV) and human immunodeficiency virus (HIV) in women coinfected with HSV and HIV
Journal of Clinical Microbiology, Volume 44, No. 2, Year 2006
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Description
The accuracy and usefulness of laboratory-developed real-time PCR procedures using a Light Cycler instrument (Roche Diagnostics) for detecting and quantifying human immunodeficiency virus type 1 (HIV-1) RNA and DNA as well as herpes simplex virus type 1 (HSV-1)/HSV-2 DNA in cervicovaginal secretions from women coinfected with HIV and HSV were evaluated. For HIV-1, the use of the NEC152 and NEC131 primer set and the NEC-LTR probe in the long terminal repeat gene allowed us to detect accurately the majority of HIV-1 subtypes of group M circulating in sub-Saharan Africa, including subtypes A, B, C, D, and G as well as circulating recombinant forms 02 and 11. The detection threshold of real-time PCR for HIV in cervicovaginal lavage samples was 5 copies per assay for both RNA and DNA; the intra- and interassay coefficients of variation of C T values were 1.30% and 0.69% (HIV-1 RNA) and 1.84% and 0.67% (HIV-1 DNA), respectively. Real-time PCR for HSV using primers and probe targeting the HSV DNA polymerase gene allowed both detection and quantification of HSV DNA and also differentiation between HSV-1 and HSV-2 genotypes. The detection threshold of real-time PCR for HSV was 5 copies per assay; the intra- and interassay coefficients of variation of CT values were 0.96% and 1.49%, respectively. Both manual and automated silica-based procedures were appropriate for combined extraction of HIV and HSV genomes from female genital secretions. Taken together, these findings indicate that real-time PCR may be used as a unique nucleic acid amplification procedure to detect and quantify HIV and HSV genomes in cervicovaginal secretions and thus to assess at reduced costs the genital shedding of both viruses in women included in intervention studies. Copyright © 2006, American Society for Microbiology. All Rights Reserved.
Authors & Co-Authors
Le-Goff, Jérôme
Unknown Affiliation
Bouhlal, Hicham
Unknown Affiliation
Grésenguet, Gérard
Unknown Affiliation
Weiss, Helen Anne
Unknown Affiliation
Khonde, Nzambi
Unknown Affiliation
Hocini, Hakim
Unknown Affiliation
Désiré, Nathalie
Unknown Affiliation
Si-Mohamed, Ali
Unknown Affiliation
de Dieu Longo, Jean
Unknown Affiliation
Chemin, Cécile
Unknown Affiliation
Frost, Eric Harold E.
Unknown Affiliation
Pépin, Jacques
Unknown Affiliation
Malkin, Jean Élie
Unknown Affiliation
Mayaud, Philippe C.
Unknown Affiliation
Bélec, Laurent
Unknown Affiliation
Statistics
Citations: 60
Authors: 15
Affiliations: 8
Identifiers
Doi:
10.1128/JCM.44.2.423-432.2006
ISSN:
00951137
Research Areas
Genetics And Genomics
Infectious Diseases
Sexual And Reproductive Health
Study Design
Randomised Control Trial
Participants Gender
Female