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Development of a LUX real-time PCR for the detection and quantification of human herpesvirus 7

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Human herpesvirus 7 is a highly seroprevalent -herpesvirus that, following primary infection, remains latent in CD4+ T cells and determines a persistent rather than a latent infection in various tissues and organs, including the lung and skin. This paper describes the development of an in-house light upon extension real-time PCR assay for quantification of human herpesvirus 7 DNA in clinical samples. The efficiency, sensitivity, specificity, inter- and intra-assay variability, and dynamic range have been determined. Subsequently, the assay has been validated by evaluating the human herpesvirus 7 load in bronchoalveolar lavages and skin specimens, chosen as 2 persistency sites, from healthy and pathological individuals. The real-time PCR assay developed in this study could be a useful tool to detect and quantify human herpesvirus 7 DNA in different clinical specimens to elucidate its epidemiological and pathogenic roles.

L’herpèsvirus-7 humain est un -herpèsvirus à séroprévalence élevée qui, suite à une infection primaire, demeure latent dans les lymphocytes T CD4+ et établit une infection persistante plutôt que latente dans différents tissus et organes, notamment le poumon et la peau. Cet article décrit le développement d’un essai en PCR en temps réel par la technologie LUX (Light Upon eXtension) pour quantifier l’ADN de l’herpèsvirus-7 humain dans des échantillons cliniques. L’efficacité, la sensibilité, la spécificité, la variabilité inter-essai et intra-essai ainsi que la gamme dynamique de l’essai ont été déterminées. Subséquemment, cet essai a été validé en évaluant la charge en HHV-7 de lavages broncho-alvéolaires et de spécimens de peau choisis comme sites d’infection persistante provenant d’individus sains ou avec pathologie. L’essai PCR en temps réel développé dans cette étude pourrait être un outil utile pour détecter et quantifier l’ADN de l’herpèsvirus-7 dans différents spécimens cliniques afin d’élucider ses caractéristiques épidémiologiques et son rôle dans la pathogénie de l’infection.

Document Type: Research Article

Publication date: 2009-03-01

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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