摘要
目的 研究一种快速、特异的检测大肠埃希菌 (以下称大肠杆菌 )O15 7:H7的复合PCR方法。方法 选用针对大肠杆菌O15 7:H7志贺样毒素Ⅰ、Ⅱ (SLT -Ⅰ、SLT -Ⅱ )和溶血素 (Hly)基因的三对引物 ,在同一扩增体系中进行PCR ,检测12株不同来源的O15 7:H7大肠杆菌及其它致病性大肠杆菌及沙门菌、志贺菌 15株。结果 除质粒缺失株 (933)外 ,其余 11株O15 7:H7大肠杆菌均在 36 1bp处有溶血素基因产物出现 ;而 12株O15 7:H7大肠杆菌扩增后的两条志贺样毒素基因产物存在差异 ,其中 6株在 2 10bp、484bp处出现两条相应产物 ,3株仅有 2 10bp一条SLT -Ⅰ基因产物 ,另 3株仅有 484bp的SLT-Ⅱ基因产物。其它致病性大肠杆菌及沙门菌、志贺菌的扩增结果均为阴性。结论 本复合PCR方法具有较高的特异性 ,可迅速、有效地将O15 7:H7大肠杆菌与其它常见致病性大肠杆菌及沙门菌、志贺菌相鉴别 ,通过进一步研究有望应用于临床大肠杆菌O15
Aim To develop a method of multiplex PCR to detect E.coli O157:H7 rapidly and specifically.Method Three pair of primers based on the shiga-lide toxin I and II(SLT-I、SLT-II) and hly gene sequences of E.coli O157:H7 different sources and other 15 strains non-O157:H7 E.coli coming from were selected for and amplification system,12 strains of E.coli O157:H7 E.coli were detected by the multiplex PCR.Results All 12 strains of E.coli O157:H7 but strain 933 losing hemolysin plasmid produced a 361bp fragment on the basis of the hlyA gene sequence,but the recults of the amplification of shiga-like toxin gene products of 12 strains E.coli O157:H7 were different,among them the PCR products of 6 strains were two fragments (210bp and 484dp) specific for the SLT-I and SLT-II gene sequence respectively ,the amplified product of 3 strains was a 210bp fragment from SLT-I gene,the product of the other 3 strains was a 484dp fragment from SLT-II gene.The PCR results of all other pathogenic E.coli and Shigella,Salmonella were negative.Conclusion This multiplex PCR method has high specificity,it offers a rapid and efficient means to distinct the E.coli O157:H7 form the other pathogenic E.coli and Shigella,Salmonella.Through further study this PCR assay would become an auxiliary method to diagnose the E.coli O157:H7 infection in clinic.
出处
《中国人兽共患病杂志》
CSCD
北大核心
2001年第3期80-82,共3页
Chinese Journal of Zoonoses
