中国草食动物科学2012，Vol.32Issue(4)：50-53,4.
肠出血性大肠杆菌O157:H7EMA-PCR检测技术的建立
Method to Detect Enterohemorrhagic Escherichia Coli O157:H7 by EMA-PCR
摘要
Abstract
Enterohemorrhagic Esckerichia coli 0157:H7 is one important zoonosis infectious disease pathogens. In order to establish a specific, sensitive new detection technology for 0l57:H7,the specific primers were designed according to the 0157 antigen gene (rfbE gene) as a template, the bacteria was treated with EMA, the bacterial lysates was prepared through the boiling water bath method,and PCR conditions were optimized,and a rapid and effective detection method of EMA-PCR for live 0157:H7 was established. The results showed that the 495 bp amplicon was specifically detected using the EMA-PCR in positive strains CVCC248 and two clinical isolates cd0912 and cd0803. The detection sensitivity was up to 12 CFU/mL By EMA treatment,the target fragment was amplified from the DNA of mixed suspension containing l%~100% live O157:H7 CVCC248. Therefore,the EMA-PCR method for 0157:H7 was successfully established. This method avoided the false positive test result caused by the sample containing dead bacteria,and improved the accuracy and authenticity of the detection in comparing with the conventional PCR. The EMA-PCR method for 0157:H7 provides a new approach in the clinical diagnosis,it has important practical value and good prospect.关键词
肠出血性大肠杆菌/rfbE基因/EMA-PCR/死细菌/活细菌Key words
Enterohemorrhagic Escherichia coli/ rfbE gene/ EMA-PCR/ dead bacteria/ live bacteria分类
农业科技引用本文复制引用
赵素君,谢晶,曹冶,李江凌,王秋实,叶勇刚,罗丹丹,叶健强,廖党金..肠出血性大肠杆菌O157:H7EMA-PCR检测技术的建立[J].中国草食动物科学,2012,32(4):50-53,4.基金项目
国家科技支撑计划项目(2006BAD04A17、2012BAD12B03) （2006BAD04A17、2012BAD12B03）
四川省科技支撑计划项目(2010NZ0106) （2010NZ0106）
国家奶牛产业技术体系岗位科学家项目 （）
四川省生猪现代产业体系项目 （）
