Establishment and Application of a Real-Time Quantitative PCR Assay for Detection of Campylobacter Jejuni in Human Feces
10.3969/j.issn.1671-7414.2014.05.021
- VernacularTitle:实时荧光定量PCR检测人粪便中空肠弯曲杆菌方法的建立与评价
- Author:
Yunpeng CHEN
;
Wen LIN
;
Jianping DENG
- Publication Type:Journal Article
- Keywords:
real-time quantitative PCR;
campylobacter jejuni;
human feces
- From:
Journal of Modern Laboratory Medicine
2014;(5):85-88
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a real-time quantitative PCR assay for the identification of Campylobacter jejuni in fecal samples.Methods Specific primers of the PCR were designed according to the conserved sequences of Campylobacterjeju-ni,and the real-time quantitative PCR assay was established.150 cases of fecal samples were tested by both culture and PCR methods.With the culture testing results as the reference standard,the sensitivity,specificity,accuracy and repetition of the real-time quantitative PCR were validated.Kappa test was used to estimate the difference between the two detection meth-ods.Results The standard carve of the real-time quantitative PCR assay fitted the equationY=-3.51Log(X)+37.09 (R2=0.996)well.The sensitivity,specificity,and accuracy of the established method were 92.4%,95.8% and 94%,respective-ly.The theoretical detection limit of the PCR method was 102 CFU/ml,and its reproducibility was good (CV<5%).Statisti-cal analysis demonstrated that the results of the two methods were consistent,and the consistent strength was very strong (Kappa=0.88,P<0.05).Conclusion The established real-time PCR method can assay the Campylobacterjejuni in human fecal samples rapidly and accurately.
