title: Multiplex real-time PCR for the simultaneous detection of herpes simplex virus, human herpesvirus 6, and human herpesvirus 7.
abstract: A simultaneous detection system to quantify HSV, HHV-6, and HHV-7 DNA via multiplex real-time PCR using different fluorochromes was developed. The minimum quantitative level established via this multiplex assay was four copies per reaction for HSV type 1, four copies for HHV-6, and three copies for HHV-7, respectively. The dynamic range encompassed at least six orders of magnitude. The system was specific and reproducible even in the presence of large amounts of other viral DNA. We then applied this multiplex real-time PCR assay to 105 CSF specimens obtained from subjects less than 15 years old in whom a diagnosis of viral encephalitis/encephalopathy was suspected on clinical grounds. The detection rate for each viral DNA was 6.7% for HSV, 9.5% for HHV-6, and 1.9% for HHV-7. These results indicate that our system is reliable and may be useful for the rapid diagnosis of viral encephalitis/encephalopathy.
authors.affiliation: Deparmtent of Virology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Kaoru
authors.initials: K
authors.lastname: Wada
authors.forename: Sachiko
authors.initials: S
authors.lastname: Mizoguchi
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Ito
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Kawada
authors.forename: Yohei
authors.initials: Y
authors.lastname: Yamauchi
authors.forename: Tsuneo
authors.initials: T
authors.lastname: Morishima
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Nishiyama
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kimura
mesh_terms: D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D018792:Encephalitis, Viral / Q000175:diagnosis / Q000821:virology; D005260:Female; D006561:Herpes Simplex / Q000175:diagnosis / Q000821:virology; D015654:Herpesvirus 6, Human / Q000235:genetics / Q000302:isolation & purification*; D016199:Herpesvirus 7, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D019349:Roseolovirus Infections / Q000175:diagnosis / Q000821:virology; D018139:Simplexvirus / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D004279:DNA, Viral