Development of a LUX real-time PCR for the detection and quantification of human herpesvirus 7.
Human herpesvirus 7 is a highly seroprevalent beta-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.
Virology Unit, Department of Public Health and Microbiology, University of Turin, Via Santena 9, Turin 10126, Italy.
Massimiliano
M
Bergallo
Cristina
C
Costa
Maria Elena
ME
Terlizzi
Francesca
F
Sidoti
Samuela
S
Margio
Sara
S
Astegiano
Renata
R
Ponti
Rossana
R
Cavallo
D001992:Bronchoalveolar Lavage Fluid / Q000821:virology; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D005456:Fluorescent Dyes; D016199:Herpesvirus 7, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D019349:Roseolovirus Infections / Q000175:diagnosis / Q000821:virology; D012680:Sensitivity and Specificity; D012867:Skin / Q000821:virology
D017931:DNA Primers; D004279:DNA, Viral; D005456:Fluorescent Dyes