聚合酶链反应法和分离培养法筛检小肠结肠炎耶尔森菌方法的比较研究
Comparison of Yersinia enterocolitica detection by PCR and conventional culturing
-
摘要: 目的 比较针对小肠结肠炎耶尔森菌的两种不同检测方法,即ail基因联合foxA基因的聚合酶链反应(polymerase chain reaction,PCR)检测和分离培养法的差异。 方法 从生猪咽拭子和回盲部肠内容物标本中提取细菌基因组DNA并进行小肠结肠炎耶尔森菌保守基因foxA和粘附侵袭位点基因ail的PCR检测;菌株分离培养按常规方法进行。将两种方法的检出结果进行统计分析,以判定哪种更有优势。 结果 700份标本中,小肠结肠炎耶尔森菌特异性基因PCR检测阳性402份,阳性率57.43%;小肠结肠炎耶尔森菌分离培养阳性278份,阳性率39.71%。在PCR检测阳性的402份标本中,分离小肠结肠炎耶尔森菌271株,分离阳性率为67.41%;PCR检测阴性的298份标本中,分离小肠结肠炎耶尔森菌7株,分离阳性率仅为2.35%。 结论 本次实验证实,检测来源于生猪标本中的小肠结肠炎耶尔森菌,采用foxA和ail基因联合的PCR检测法,检出率明显高于小肠结肠炎耶尔森菌分离培养法(P0.05)。Abstract: Objective To compare the performance of polymerase chain reaction (PCR) and conventional culturing in the detection of Yersinia enterocolitica. Methods The swine throat swabs and intestine content samples were collected to extract bacterium genomic DNA and detect specific genes ail and foxA of Y. enterocolitica with PCR amplification, and to isolate Y. enterocolitica by conventional culturing. Results Among the 700 samples collected, 402 (57.43%) were positive in PCR amplifications and 278 (39.71%) were positive in conventional culturing. There were 271 Y. enterocolitica strains isolated from the 402 PCR positive samples by conventional culturing (67.41%) and 7 Yersinia strains isolated from the 298 PCR negative samples by conventional culturing (2.35%). Conclusion The sensitivity of PCR amplification of ail and foxA is significant higher than that of conventional culturing in detecting Y. enterocolitica in swine throat swabs and intestine content samples (P0.05).
下载: