牛乳中大肠杆菌O157:H7 PCR检测方法的建立

Establishment of PCR Method for Detection of Escherichia coli O157:H7 in Milk

为了建立牛乳中大肠杆菌O157∶H7的PCR快速检测方法,试验对大肠杆菌O157∶H7的rfbE基因序列进行分析,设计1对特异性引物,对大肠杆菌O157∶H7的DNA进行PCR扩增,结合特异性和灵敏性试验,实现对致病性大肠杆菌O157∶H7的检测;然后将不同含量的大肠杆菌掺入到牛乳中进行实际样品的检测。结果表明,该方法的特异性和灵敏性较好,能够检测到0.1 ng/μL的DNA模板,当牛乳中含有超过100 CFU/mL的大肠杆菌时,可利用该方法进行检测。

In order to establish PCR rapid detection method on Escherichia coli（E. coli） O157：H7 of milk. The rfb E gene sequence of E. coli O157 ：H7 is analyzed in the experiment and a pair of specific primers are designed. E. coli DNA is amplified by PCR,combined with the specificity and sensitivity experiments,the detection of pathogenic E. coli O157：H7 is implemented. Then different content of E. coli are added to milk to detect the actual samples. The results show that this method had good specificity and sensitivity,can detected 0.1 ng/μL DNA template,when milk contains more than 100 CFU/m L E. coli,this method can be used for testing.