A Multiplex PCR Assay for the Detection of Pathogenic Genes of EPEC, ETEC and EIEC

• 作 者: ZHANG Tienan LI Jichang LU Che
• 机 构: College of Veterinary Medicine
• 出 处: 东北农业大学学报（英文版）
• 发表日期: 2004年期
• 关键词: enteropathogenic Escherichia coli. (EPEC) enterotoxigenic Escherichia coli. (ETE
• 基金项目: Key Item of National Technology Research Project;黑龙江省资助项目

A multiplex polymerase chain reaction (PCR) was developed to detect three pathogenic genes of enteropathogenic, enterotocigenic and enteroinvasive Escherichia coli.. In this study three different sets of oligonucleotide primer were simultaneously used, and in this way, specific fragments of 880, 600, 150 bp for EPEC eaeA,EIEC ipaH and ETEC ST genes were amplified, respectively. The best condition of the multiplex PCR was: after an initial heat denaturation step at 95℃ for 5 min, followed by 30 cycles of denaturation at 94 ℃ for 40 s, primer annealing at 51.3 ℃ for 40 s and extension at 72 ℃ for 1 min, final extension at 72 ℃ for 10 min. The detection limit of tively. It may be a good way for the detection and identification of Diarrhea-causing E. coli..