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 equi...

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Bibliographic Details
Published in:Microbiology and immunology 2014-01, Vol.58 (1), p.68-71
Main Authors: Niwa, Shoichi, Tsukagoshi, Hiroyuki, Ishioka, Taisei, Sasaki, Yoshiko, Yoshizumi, Masakazu, Morita, Yukio, Kimura, Hirokazu, Kozawa, Kunihisa
Format: Article
Language:English
Subjects:
Caliciviridae Infections - diagnosis
Caliciviridae Infections - virology
Feces
Genes
Humans
Multiplex Polymerase Chain Reaction
multiplex real-time polymerase chain reaction
Norovirus
Norovirus - genetics
Real-Time Polymerase Chain Reaction
Reproducibility of Results
sapovirus
Sapovirus - genetics
Sensitivity and Specificity
Viral Load - methods
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Summary: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 102–107 copies/assay using plasmid DNA standards. The limit of detection was 102 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.
ISSN:0385-5600
1348-0421
DOI:10.1111/1348-0421.12107