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目的通过对婴幼儿腹泻病例粪便标本进行致病性大肠埃希菌的分离鉴定及毒力基因检测,从分子生物学方面研究致病性大肠埃希菌引起婴幼儿腹泻的分布,以降低婴幼儿致病性大肠埃希菌的感染率。方法收集江西省儿童医院2011-2012年婴幼儿腹泻病例357份粪便标本,按照致病性大肠埃希菌分离鉴定程序用EC肉汤增菌过夜,增菌后用麦康凯琼脂培养基分离培养,可疑菌用API 20E生化鉴定出大肠埃希菌,然后用聚合酶链反应(PCR)检测致病性大肠埃希菌相应的毒力基因。结果 357份腹泻患者粪便标本中,检出37份携带毒力基因的致病性大肠埃希菌,检出率为10.36%,携带毒力基因的肠聚集性大肠埃希菌(EAggEC)、肠道致病性大肠埃希菌(EPEC)、肠道侵入性大肠埃希菌(EIEC)、肠道出血性大肠埃希菌(EHEC)检出率分别为8.4%、0.56%、1.12%、0.28%,产肠毒素大肠埃希菌(ETEC)未检出。结论婴幼儿腹泻病例中致病性大肠埃希菌主要由EaggEC引起,其次为EIEC,进一步证实对致泻大肠埃希菌进行PCR毒力基因检测具有重要临床意义。
Objective To investigate the distribution of diarrhea in infants and young children caused by pathogenic Escherichia coli in molecular biology to isolate and identify pathogenic Escherichia coli in stool samples from infants and young children, Pathogenic Escherichia coli infection rate. Methods 357 stool samples from infants and young children with diarrhea in Jiangxi Children’s Hospital from 2011 to 2012 were collected and stained with EC broth according to the procedure of isolation and identification of pathogenic Escherichia coli. After enrichment, the cells were separated and cultured with MacConkey agar medium, Escherichia coli was identified biochemically by API 20E, and the corresponding virulence genes of pathogenic Escherichia coli were detected by polymerase chain reaction (PCR). Results Among 357 diarrhea patients, 37 pathogenic Escherichia coli strains carrying virulence genes were detected, the detection rate was 10.36%, intestinal aggregated Escherichia coli (EAggEC) carrying virulence genes, The detection rates of EPEC, EIEC and EHEC were 8.4%, 0.56%, 1.12%, 0.28 %, Enterotoxigenic Escherichia coli (ETEC) was not detected. Conclusions Pathogenic Escherichia coli is mainly caused by EaggEC in infantile diarrhea cases, followed by EIEC. It is further confirmed that it is of clinical significance to detect PCR virulence genes in Escherichia coli diarrhea.