SYBR green I real-time polymerase chain reaction for detection of Norovirus II in the shellfish.
- Author:
Xuemei MO
1
;
Dongwei GAO
Author Information
1. Institute of Genome Medicine, Jinan University, Guangzhou 510632, China. mxm200101@163.com
- Publication Type:Journal Article
- MeSH:
Animals;
Bivalvia;
virology;
Norovirus;
isolation & purification;
Organic Chemicals;
RNA, Viral;
analysis;
isolation & purification;
Reverse Transcriptase Polymerase Chain Reaction;
methods;
Sensitivity and Specificity;
Shellfish;
virology;
Species Specificity
- From:
Chinese Journal of Biotechnology
2010;26(6):817-822
- CountryChina
- Language:Chinese
-
Abstract:
We set up an SYBR Green I real-time RT-PCR method for the detection of genogroup II Norovirus, and this method's primers were encompassed the conservative region of Norovirus II. The limit of the detection was 10(2) copies. The standard curve's linear range was 10(2)-10(6) copies, correlation coefficient was 0.9952, the slope was -2.982, and the intercept was 35.84. This method possessed specificity for genogroup II Norovirus, without any cross-reaction with rotavirus, adenovirus, hepatitis A virus or astrovirus. The coefficients of variation (CV) of the C(t) values of the standard plasmid were 0.95%-1.69% (n = 5) in intra-assay and 0.87%-1.24% (n = 3) in inter-assay. We used this method to detect 30 shellfish samples, and found 3 samples were positive. This method is sensitive, specific and reliable for Norovirus II. It can be used to detect the Norovirus II in the shellfish rapidly.
