Detection of Escherichia coli O157:H7 by SYBR Green I real-time ramification amplification method

  Objective   To establish SYBRGreen I real-time ramification amplification method(RAM)fordetection of Escherichia coli O157:H7.

  Methods   The bacteria were diluted in 100-fold serial with saline from 107 to 10 bacteria/Ll, and detected by rea-l time RAMfordetermining the sensitivity.Clinical isolates were detected forfurtherstudy.

  Results   The lowest numberof targets detected by rea-l time RAMwas 10 bacteria/Ll, and the time of appearance of detectable signal was depended on the target concentration.Three clinical isolates of E.coli O157:H7 were positive forshig a tox in gene, while nonpathogenic E.coli were negative.

  Conclusion   SYBRGreen Ireal-time RAMis rapid, accurate, simple, real-time environ-mental protecting, and could be another DNA amplification method to detect E.coli O157:H7.