Al-Ajmi D, Padmanabha J, Denman S E, Gilbert R A, Al Jassim R A M, McSweeney C S
School of Animal Studies, University of Queensland, Gatton, Qld, Australia.
Lett Appl Microbiol. 2006 Apr;42(4):386-91. doi: 10.1111/j.1472-765X.2005.01850.x.
Combinations of PCR primer sets were evaluated to establish a multiplex PCR method to specifically detect Escherichia coli O157:H7 genes in bovine faecal samples.
A multiplex PCR method combining three primer sets for the E. coli O157:H7 genes rfbE, uidA and E. coli H7 fliC was developed and tested for sensitivity and specificity with pure cultures of 27 E. coli serotype O157 strains, 88 non-O157 E. coli strains, predominantly bovine in origin and five bacterial strains other than E. coli. The PCR method was very specific in the detection of E. coli O157:H7 and O157:H- strains, and the detection limit in seeded bovine faecal samples was <10 CFU g(-1) faeces, following an 18-h enrichment at 37 degrees C, and could be performed using crude DNA extracts as template.
A new multiplex PCR method was developed to detect E. coli O157:H7 and O157:H-, and was shown to be highly specific and sensitive for these strains both in pure culture and in crude DNA extracts prepared from inoculated bovine faecal samples.
This new multiplex PCR method is suitable for the rapid detection of E. coli O157:H7 and O157:H- genes in ruminant faecal samples.
评估聚合酶链反应（PCR）引物组的组合，以建立一种多重PCR方法，用于特异性检测牛粪便样本中的大肠杆菌O157:H7基因。
开发了一种多重PCR方法，该方法结合了用于大肠杆菌O157:H7基因rfbE、uidA和大肠杆菌H7 fliC的三组引物，并对27株大肠杆菌O157血清型菌株、88株非O157大肠杆菌菌株（主要来源于牛）以及5株非大肠杆菌菌株的纯培养物进行了敏感性和特异性测试。该PCR方法在检测大肠杆菌O157:H7和O157:H-菌株时具有高度特异性，在接种的牛粪便样本中，经37℃ 18小时富集后，检测限<10 CFU g(-1)粪便，并且可以使用粗DNA提取物作为模板进行检测。
开发了一种新的多重PCR方法来检测大肠杆菌O157:H7和O157:H-，并证明该方法在纯培养物以及从接种的牛粪便样本制备的粗DNA提取物中对这些菌株具有高度特异性和敏感性。
这种新的多重PCR方法适用于快速检测反刍动物粪便样本中的大肠杆菌O157:H7和O157:H-基因。
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