Publication: Evaluation of real-time RT-PCR assays for detection and quantification of norovirus genogroups I and II
Issued Date
2017-04-01
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ISSN
1995820X
16740769
16740769
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2-s2.0-85013480694
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Mahidol University
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SCOPUS
Bibliographic Citation
Virologica Sinica. Vol.32, No.2 (2017), 139-146
Suggested Citation
Kitwadee Rupprom, Porntip Chavalitshewinkoon-Petmitr, Pornphan Diraphat, Leera Kittigul Evaluation of real-time RT-PCR assays for detection and quantification of norovirus genogroups I and II. Virologica Sinica. Vol.32, No.2 (2017), 139-146. doi:10.1007/s12250-016-3863-9 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/41912
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Title
Evaluation of real-time RT-PCR assays for detection and quantification of norovirus genogroups I and II
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Abstract
© 2017, Wuhan Institute of Virology, CAS and Springer Science+Business Media Singapore. Noroviruses are the leading cause of acute gastroenteritis in humans. Real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) is a promising molecular method for the detection of noroviruses. In this study, the performance of three TaqMan real-time RT-PCR assays was assessed, which were one commercially available real-time RT-PCR kit (assay A: Norovirus Real Time RT-PCR kit) and two in-house real-time RT-PCR assays (assay B: LightCycler RNA Master Hybprobe and assay C: RealTime ready RNA Virus Master). Assays A and B showed higher sensitivity than assay C for norovirus GI, while they all had the same sensitivity (103 DNA copies/mL) for GII DNA standard controls. Assay B had the highest efficiency for both genogroups. No cross-reactivity was observed among GI and GII noroviruses, rotavirus, hepatitis A virus, and poliovirus. The detection rates of these assays in GI and GII norovirus-positive fecal samples were not significantly different. However, the mean quantification cycle (Cq) value of assay B for GII was lower than assays A and C with statistical significance (P-value, 0.000). All three real-time RT-PCR assays could detect a variety of noroviruses including GI.2, GII.2, GII.3, GII.4, GII.6, GII.12, GII.17, and GII.21. This study suggests assay B as a suitable assay for the detection and quantification of noroviruses GI and GII due to good analytical sensitivity and higher performance to amplify norovirus on DNA standard controls and clinical samples. [Figure not available: see fulltext.].