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大肠杆菌O118 O抗原基因簇中血清群特异性基因的DNA测序与鉴定,以及大肠杆菌O118和O151 O抗原簇DNA序列的抗原多样性及微小变异的证明。

DNA sequencing and identification of serogroup-specific genes in the Escherichia coli O118 O antigen gene cluster and demonstration of antigenic diversity but only minor variation in DNA sequence of the O antigen clusters of E. coli O118 and O151.

作者信息

Liu Yanhong, Fratamico Pina, Debroy Chitrita, Bumbaugh Alyssa C, Allen John W

机构信息

U.S. Department of Agriculture, Agricultural Research Service, Eastern Regional Research Center, Wyndmoor, Pennsylvania 19038, USA.

出版信息

Foodborne Pathog Dis. 2008 Aug;5(4):449-57. doi: 10.1089/fpd.2008.0096.

Abstract

The DNA sequence of the O antigen gene cluster of an Escherichia coli serogroup O118 strain was determined, and 13 open reading frames (ORFs) were identified, encoding genes required for O antigen sugar biosynthesis, transfer, and processing. Polymerase chain reaction (PCR) assays targeting the wzx (O antigen flippase) and wzy (O antigen polymerase) genes in the O antigen gene cluster of E. coli O118 were designed for identification of these serogroups. Specificity testing using strains belonging to E. coli O118 isolated from various sources, representative strains of 167 other E. coli O serogroups, and 20 non-E. coli bacteria revealed that the PCR assays were specific for E. coli O118. Thus, the PCR assays can be used for rapid identification of E. coli O118 as an alterative to typing using antisera. However, the PCR assays targeting the E. coli O118 wzx and wzy genes were also positive using E. coli serogroup O151 DNA. Therefore, the sequence of the O antigen gene cluster of E. coli O151 was determined, and it was very similar to that of E. coli O118, with only three nucleotide differences. Although the lipopolysaccharide profiles of O118 and O151 showed differences, multilocus sequence typing of E. coli O118 and O151 strains only revealed minor variation at the nucleotide level. Since E. coli O118 strains are more frequently isolated from humans, animals, and the environment than E. coli O151, serogroup O151 may likely be a minor variant of E. coli O118. Further studies are needed to elucidate this possibility.

摘要

测定了一株大肠杆菌O118血清群菌株的O抗原基因簇的DNA序列,鉴定出13个开放阅读框(ORF),它们编码O抗原糖生物合成、转移和加工所需的基因。设计了针对大肠杆菌O118的O抗原基因簇中的wzx(O抗原翻转酶)和wzy(O抗原聚合酶)基因的聚合酶链反应(PCR)检测方法,用于这些血清群的鉴定。使用从各种来源分离的大肠杆菌O118菌株、其他167种大肠杆菌O血清群的代表性菌株以及20种非大肠杆菌细菌进行特异性测试,结果表明该PCR检测方法对大肠杆菌O118具有特异性。因此,该PCR检测方法可作为使用抗血清分型的替代方法,用于快速鉴定大肠杆菌O118。然而,使用大肠杆菌O151血清群的DNA进行检测时,针对大肠杆菌O118 wzx和wzy基因的PCR检测也呈阳性。因此,测定了大肠杆菌O151的O抗原基因簇序列,发现它与大肠杆菌O118的序列非常相似,仅存在三个核苷酸差异。尽管O118和O151的脂多糖谱存在差异,但大肠杆菌O118和O151菌株的多位点序列分型仅在核苷酸水平上显示出微小差异。由于从人类、动物和环境中分离出的大肠杆菌O118菌株比大肠杆菌O151更频繁,血清群O151可能是大肠杆菌O118的一个小变体。需要进一步研究以阐明这种可能性。

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