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Triplex real‐time polymerase chain reaction assay for detection and quantification of norovirus ( GI and GII ) and sapovirus
Author(s) -
Niwa Shoichi,
Tsukagoshi Hiroyuki,
Ishioka Taisei,
Sasaki Yoshiko,
Yoshizumi Masakazu,
Morita Yukio,
Kimura Hirokazu,
Kozawa Kunihisa
Publication year - 2014
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/1348-0421.12107
Subject(s) - sapovirus , norovirus , real time polymerase chain reaction , biology , detection limit , polymerase chain reaction , virology , microbiology and biotechnology , plasmid , acute gastroenteritis , dna , chromatography , chemistry , genetics , virus , gene
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.