Establishment of a PCR method for detection of Enterobacter sakazakii

Objective To establish a PCR method for detection of Enterobacter sakazakii.Methods DNA was extracted by thermal denaturalization method and phenol-chloroform method respectively.A pair of oligonucleotide primers were designed with ompA gene to detect Enterobacter sakazaki.In addition,PCR conditions were optimized,and the specificity and sensitivity of PCR were determined.Results The optimized PCR condition were:the concentration of Mg2+ was 2.0mmol/l,the anealing temperature was 60℃.The predicted specific DNA amplcation bands of 469bp were obtained and the sensitivity of thermal denaturalization method and phenol-chloroform method revealed that as little as 105CFU/ml and 103CFU/ml of products could be detected,respcetively.Conclusion The method is specific,sensitive,and rapid.It is useful for the rapid detection of Enterobacter sakazakii.