Development of hepatitis C virus by fluorescent real-time reverse transcription loop-mediated isothermal amplification method
10.3760/cma.j.issn.0254-5101.2011.06.020
- VernacularTitle:荧光定量环介导逆转录等温扩增技术检测丙型肝炎病毒反应体系的建立
- Author:
Yongle ZHANG
;
Shourong LIU
;
Jing YANG
- Publication Type:Journal Article
- Keywords:
Fluorescence quantitation;
Reverse loop-mediated isothermal amplification;
Hepatitis C virus;
Real-time PCR
- From:
Chinese Journal of Microbiology and Immunology
2011;31(6):564-566
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a rapid, sensitive, and specific quantitative method to detect hepatitis C virus. Methods A primer set targeting HCV 5'UTR was designed. The isothermal amplification was performed by the Bst DNA polymerase and AMV reverse transcriptase, under the temperature of 60℃ for 60 min. The signal was monitored by SYBR Green Ⅰ. Results One hundred and twenty positive serum samples, confirmed by the real-time PCR. All were detected by the isothermal amplification, while 110 healthy subjects' samples were negative by the both methods. The lower detect limit was determined to 10 IU/ml HCV-RNA, by the assay on serial dilutions of the quality control standards obtained from clinical investigation center of MOH. Conclusion A real time reverse loop-mediated isothermal amplification method was developed to detect HCV, with the characteristic of rapidity, high sensitivity and specificity.
