Abstract：
Escherichia albertii is an emerging enteropathogen. Several foodborne outbreaks of E. albertii have been reported in Japan; however, foods associated with most outbreaks remain unidentified. Therefore, polymerase chain reaction (PCR) assays detecting E. albertii specifically and sensitively are required. Primers and probe for real-time PCR assays targeting E. albertii-specific gene (EA-rtPCR) was designed. With 74 strains, including 43 E. albertii strains and several of its close relatives, EA-rtPCR specifically amplified E. albertii; therefore, the sensitivity of EA-rtPCR was then evaluated. The detection limits were 2.8 and 2.0-3.2 log colony-forming unit (CFU)/mL for E. albertii culture and enriched chicken culture inoculated with the pathogen, respectively. In addition, E. albertii was detected from 25 g of chicken meat inoculated with 0.1 log CFU of the pathogen by EA-rtPCR. The detection of E. albertii from chicken meat by EA-rtPCR was also evaluated by comparing with the nested-PCR assay, and 28 retail chicken meat and 193 dissected body parts from 21 chicken carcass were tested. One and three chicken meat were positive in the nested-PCR assay and EA-rtPCR, respectively. Fourteen carcasses had at least one body part that was positive for EA-rtPCR, and 36 and 48 samples were positive for the nested-PCR assay and EA-rtPCR, respectively. A total of 37 strains of E. albertii were isolated from seven PCR-positive samples obtained from six chicken carcass. All E. albertii isolates harbored eae gene, and were classified as E. albertii O-genotype (EAOg)3 or EAOg4 by EAO-genotyping. The EA-rtPCR developed in this study has potential to improve E. albertii detection in food and advance research on E. albertii infection.
摘要：
艾氏大肠杆菌是一种新兴的肠病原体。日本已经报道了几起食源性的阿伯蒂伊病毒暴发；然而，与大多数疫情相关的食物仍未被确认。因此,需要特异性和灵敏地检测阿氏大肠杆菌的聚合酶链反应（PCR）测定法。设计了用于靶向阿氏大肠杆菌特异性基因(EA-rtPCR)的实时PCR测定的引物和探针。74株，包括43株艾伯蒂埃及其近亲，EA-rtPCR特异性扩增阿氏大肠杆菌；因此，然后评估EA-rtPCR的敏感性。艾氏大肠杆菌培养物和接种病原体的富集鸡培养物的检测限为2.8和2.0-3.2对数菌落形成单位（CFU）/mL，分别。此外,通过EA-rtPCR从接种有0.1logCFU的病原体的25g鸡肉中检测到艾氏大肠杆菌。通过与巢式PCR方法的比较，还评估了EA-rtPCR检测鸡肉中的白葡萄酒。并测试了28个零售鸡肉和来自21个鸡肉尸体的193个解剖身体部位。1个和3个鸡肉在巢式PCR和EA-rtPCR中呈阳性，分别。14具尸体至少有一个身体部位对EA-rtPCR呈阳性，36和48个样本的巢式PCR和EA-rtPCR检测呈阳性，分别。从从6只鸡car体中获得的7个PCR阳性样品中分离出37株艾氏大肠杆菌。所有艾氏大肠杆菌分离株都带有eae基因，并通过EAO基因分型分类为艾氏O基因型（EAOg）3或EAOg4。本研究中开发的EA-rtPCR有可能改善食品中的艾贝氏杆菌检测并促进艾贝氏杆菌感染的研究。
