Real-time PCR development for identification of Campylobacter coli from stool specimen
10.3969/cjz.j.issn.1002-2694.2012.10.001
- VernacularTitle:结肠弯曲菌Real-time PCR检测方法的建立
- Author:
Bo QIAO
;
Xuebin XU
;
Yixin GU
;
Guodong LIU
;
Fei ZHAO
;
Lihua HE
;
Jianzhong ZHANG
;
Maojun ZHANG
- Publication Type:Journal Article
- Keywords:
Campylobacter coli;
real-time PCR;
ceuE
- From:
Chinese Journal of Zoonoses
2012;(10):969-972
- CountryChina
- Language:Chinese
-
Abstract:
The aim of this study is to develop the real-time PCR assays to detect and qualify C. coli from stool specimen. The primers and probe were designed based on the specific sequence of ceuE gene in C. coli using Primer 5.0 and Vector NTI Suite 6.0 e. The PCR assay was optimized with the reference C. coli strains. The standard curve based on the dilutions of genomic DNA showed a linear relationship between log CFU/mL and threshold cycles. The detectable limitation was 5.62 CFU/mL by using purified DNA from bacteria culture. The reproducibility of this assay was assessed by calculating the variation of the threshold cycle value and the slope from test repeats for the same strains and different strains. Our results indicate that the developed assay has high sensitivity and specificity for identification of C. coli.
