Study on the use of TaqMan Real-time PCR to detect genogroup i and Ii norovirus in oysters and patients' stool samples
Sun, Y.-P.; Cheng, M.; Song, S.-L.; Zhang, X.-H.; Li, R.
Zhonghua Liu Xing Bing Xue Za Zhi 29(6): 594-597
2008
ISSN/ISBN: 0254-6450 PMID: 19040045 Document Number: 617004
To develop a rapid, specific and sensitive diagnostic method for quantification and typing of genogroup I and II norovirus in oyster shellfish and stool samples from patients who had eaten them. Specific primers and probe, following large scale norovirus genome consensus analysis were designed and subsequently a TaqMan based Real-time PCR assay to detect both GI and GII were established. This method showed high specificity for norovirus nucleic acid detection, and no cross-reaction among norovirus GI and GII. The limit on detection of NV genomes was 10(2) copies/microl. A total of 90 oysters and 37 stool specimens with diarrhea were tested for norovirus by conventional reverse transcriptional PCR (RT-PCR) assay as well as the TaqMan Real-time PCR, respectively. The norovirus detection rate in oysters by TaqMan PCR was significantly higher than that by conventional RT-PCR, but no differences between the two PCR methods were found when detecting the stool samples. Reliability of the Real-time PCR for norovirus detection was further confirmed by DNA sequencing of the positive samples This TaqMan Real-time PCR assay was proved to be a useful method for quantification and typing for norovirus in routine monitoring of both oyster shellfish and clinical samples. This method is recommended to be an effective diagnostic method for outbreak-associated gastroenteritis due to norovirus.