实时荧光定量PCR技术鉴别非O157:H7大肠杆菌产志贺毒素菌株(STEC)的研究

Identification of Shiga toxin producing strains (STEC) of non-O157:H7 Escherichia coli by real-time fluorescence quantitative polymerase chain reaction

本文运用实时荧光定量PCR的技术对菌株进行stx1基因、stx2基因、eaeA毒力基因检测;并对stx阳性、eaeA阳性的菌株进行O抗原基因rfbE(O157)、wzx(O26)、wbdI(O111)、ihp1(O145)、wzx(O103)检测。探究了实验室保存的94株非O157:H7大肠杆菌是否存在产志贺毒素菌株(STEC)存在;结果表明94株大肠杆菌中检出3株含有stx基因、12株含有eaeA基因;对stx和eaeA阳性菌株O抗原基因试验,检出2株含有wzx(O26)基;这2株大肠杆菌血清凝集试验结果为阳性。研究结果显示,实时荧光定量PCR技术具有特异性强,灵敏度高等特点,可用于产志贺毒素菌株(STEC)前期筛查。

In this paper,the virulence genes of stx1,Stx2 and eaeA were detected by real-time fluorescence quantitative PCR,and the O antigen genes rfbE(O157),wzx(O26),wbdI(O111),ihp1(O145),wzx(O103)were detected in stx-positive and eaeA-positive strains.The existence of Shiga toxin-producing strains(STEC)in 94 strains of non-O157:H7 E.coli preserved in laboratory was investigated.The results showed that 3 of 94 strains of E.coli contained stx gene and 12 strains contained eaeA gene.Two strains of E.coli contained wzx(O26)gene by O antigen gene test of stx and eaeA positive strains.The results of serum agglutination test of these two strains were positive.The results showed that the real-time fluorescence quantitative PCR technology could be used for pre-screening of Shiga toxin-producing strains(STEC)with the characteristics of high specificity and sensitivity.