title: Triplex real-time polymerase chain reaction assay for detection and quantification of norovirus (GI and GII) and sapovirus.
abstract: To improve detection of norovirus (NoVGI, NoVGII) and sapovirus (SaV), a simultaneous quantitative RT-PCR method was established. This triplex real-time PCR method was evaluated using a combination of optimized specific primers and probes. The performance of the developed PCR assay was equivalent to that of monoplex real-time PCR across a broad dynamic range of 10(2) -10(7) copies/assay using plasmid DNA standards. The limit of detection was 10(2) copies/assay. The quantitative value was comparable with that of monoplex real-time PCR of stool samples. Our triplex real-time PCR is useful for detection of NoV and SaV infections.
authors.affiliation: Gunma Prefectural Institute of Public Health and Environmental Sciences, 378 Kamioki-machi, Maebashi-shi, Gunma, 371-0052.
authors.forename: Shoichi
authors.initials: S
authors.lastname: Niwa
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tsukagoshi
authors.forename: Taisei
authors.initials: T
authors.lastname: Ishioka
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Sasaki
authors.forename: Masakazu
authors.initials: M
authors.lastname: Yoshizumi
authors.forename: Yukio
authors.initials: Y
authors.lastname: Morita
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Kimura
authors.forename: Kunihisa
authors.initials: K
authors.lastname: Kozawa
mesh_terms: D017250:Caliciviridae Infections / Q000175:diagnosis / Q000821:virology; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D029322:Norovirus / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D029341:Sapovirus / Q000235:genetics*; D012680:Sensitivity and Specificity; D019562:Viral Load / Q000379:methods
keywords: multiplex real-time polymerase chain reaction; norovirus; sapovirus