Development of a novel real-time polymerase chain reaction assay to detect Escherichia albertii in chicken meat.
Development of a novel real-time polymerase chain reaction assay to detect Escherichia albertii in chicken meat.
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开发一种新型实时聚合酶链反应测定法来检测鸡肉中的埃希氏菌。
DOI：
10.1089/fpd.2022.0042
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发表时间：
2022
影响因子：
2.8
通讯作者：
Yukiko Hara-Kudo.
中科院分区：
文献类型：
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作者：
Sakura Arai;Tadasuke Ooka;Mizuha Shibata;Yuhki Nagai;Yuki Tokoi;Hiromi Nagaoka;Rika Maeda;Akihiko Tsuchiya;Yuka Kojima;Kenji Ohya;Takahiro Ohnishi;Noriko Konishi;Kayoko Ohtsuka;Yukiko Hara-Kudo.
艾伯氏杆菌是一种新兴的肠道病原体。几次食源性大肠杆菌暴发。albertii已在日本报告;然而，与大多数疫情有关的食品仍未确定。因此，聚合酶链反应（PCR）检测E。要求具有特异性和敏感性。用于靶向E的实时PCR测定的引物和探针。设计了一个赤霉病菌特异性基因（EA-rtPCR）。74个菌株，包括43 E. albertiisstrains及其几个近亲，EA-rtPCR特异性地扩增了E.因此，EA-rtPCR的灵敏度进行了评价。E的检出限分别为2.8和2.0 - 3.2对数菌落形成单位（CFU）/mL。分别接种该病原体的阿尔贝提养殖和增菌鸡培养物。此外，E.用EA-rtPCR检测25 g鸡肉中的albertii，结果表明，在0.1 log CFU的接种量下，鸡肉中检出albertii。E.对28只零售鸡肉和21只鸡尸体的193个解剖部位进行了检测。巢式PCR和EA-rtPCR分别检测出1份和3份鸡肉样品。14具尸体至少有一个身体部位是阳性的EA-rtPCR，36和48个样本是阳性的巢式PCR检测和EA-rtPCR，分别。共检测到37株E.从6只鸡的7个PCR阳性样品中分离到albertii。AllE。albertiisolates harboreaegene，并归类为E.通过EAO-基因分型确定Albertio-基因型（EAOg）3或EAOg 4。本研究建立的EA-rtPCR方法具有提高E.食品中苜蓿的检测及研究进展阿尔伯蒂感染
Escherichia albertiiis an emerging enteropathogen. Several foodborne outbreaks ofE. albertiihave been reported in Japan; however, foods associated with most outbreaks remain unidentified. Therefore, polymerase chain reaction (PCR) assays detectingE. albertiispecifically and sensitively are required. Primers and probe for real-time PCR assays targetingE. albertii-specific gene (EA-rtPCR) was designed. With 74 strains, including 43E. albertiistrains and several of its close relatives, EA-rtPCR specifically amplifiedE. 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 forE. albertiiculture and enriched chicken culture inoculated with the pathogen, respectively. In addition,E. albertiiwas detected from 25 g of chicken meat inoculated with 0.1 log CFU of the pathogen by EA-rtPCR. The detection ofE. albertiifrom 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 ofE. albertiiwere isolated from seven PCR-positive samples obtained from six chicken carcass. AllE. albertiiisolates harboredeaegene, and were classified asE. albertiiO-genotype (EAOg)3 or EAOg4 by EAO-genotyping. The EA-rtPCR developed in this study has potential to improveE. albertiidetection in food and advance research onE. albertiiinfection.