Establishment and Application of Fluorescence Quantitative PCR for Enteropathogenic Escherichia coli in Rabbit
In order to develop an effective fluorescent quantitative PCR(qPCR)for detection of enteropathogenic Escherichia coli(EPEC),specific primers and probes were designed based on eaeA gene sequence,and the concentrations of primers and probes were optimized,respectively,followed by evaluation of the sensitivity,specificity and repeatability.Meanwhile,the method was used to detect clinical rabbit fecal samples and compared with the method recommended in national standard.The results showed that the detection limit of the qPCR was 2.97 copies/μL for positive plasmids,the coefficients of variation(CV)of repetition assays in intra-and inter-batch were both lower than 3%,and the number of positive clinical samples detection by qPCR was larger than that by the method recommended in national standard.In conclusion,the established method was with good specificity,high sensitivity and ideal reproducibility,and could be used to accurately detect EPEC nucleic acids in rabbit fecal samples with a good application prospect.
万方数据
维普
