Real-Time PCR Quantification of Genital Shedding of Herpes Simplex Virus (HSV) and Human Immunodeficiency Virus (HIV) in Women Coinfected with HSV and HIV
| Autor: | Cécile Chemin, Nzambi Khonde, Jacques Pépin, Nathalie Désiré, Jérôme LeGoff, Gérard Grésenguet, Ali Si-Mohamed, Hakim Hocini, Eric Frost, Helen A. Weiss, Jean-Elie Malkin, Jean De Dieu Longo, Laurent Bélec, Hicham Bouhlal, Philippe Mayaud |
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| Rok vydání: | 2006 |
| Předmět: |
Microbiology (medical)
Branched DNA Signal Amplification Assay viruses Herpesvirus 2 Human HIV Infections Cervix Uteri Biology medicine.disease_cause Polymerase Chain Reaction Herpesviridae Virus law.invention Proviruses law Virology medicine Humans Viral shedding Polymerase chain reaction Herpes Genitalis Reproducibility of Results Viral Load Molecular biology Virus Shedding Herpes simplex virus Real-time polymerase chain reaction DNA Viral Vagina HIV-1 RNA Viral Female Primer (molecular biology) Viral load |
| Zdroj: | Journal of Clinical Microbiology. 44:423-432 |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.44.2.423-432.2006 |
| Popis: | 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 C T 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. |
| Databáze: | OpenAIRE |
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