Establishment and application of a TaqMan real-time PCR assay for the detection of encephalomyo-carditis virus
10.3760/cma.j.issn.0254-5101.2013.09.013
- VernacularTitle:脑心肌炎病毒 TaqMan real-time PCR检测方法的建立及应用
- Author:
Haixia ZHANG
;
Ruofei FENG
;
Dan WANG
;
Jingjing FAN
;
Jingying XIE
;
Zhongren MA
;
Yuping FENG
- Publication Type:Journal Article
- Keywords:
Encephalomyocarditis virus ( EMCV);
Real-time PCR
- From:
Chinese Journal of Microbiology and Immunology
2013;(9):706-710
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a TaqMan real-time PCR assay for the detection of encephalo-myocarditis virus ( EMCV) .Methods Based on the conservative region of 3D gene of EMCV published in GenBank , a pair of primers and one TaqMan probe were designed and synthesized .Then a TaqMan real-time PCR assay was set up and the reactive system was optimized .The sensitivity and specificity of the assay was evaluated respectively .The TaqMan real-time PCR assay was then carried out to detect 98 randomly selected swine serum samples and the results were compared with those by using ELISA .Results The Ct value of the templates had a good linear relationship with the log starting quantity , with a correlation coefficient of 0.995.The TaqMan real-time PCR assay was only specific for EMCV and its sensitivity was 100 times higher than that of the ordinary PCR .The coincidence rate between the established assay and the ELISA assay was 98.0%in the detection of 98 blood samples.Conclusion The TaqMan real-time PCR assay for the detec-tion of EMCV was successfully established with advantages of high sensitivity and good specificity .It could be used for detection of EMCV and quantitative analysis .
