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首页> 外文期刊>Journal of Clinical Microbiology >Enzyme-linked immunosorbent assay for products of the 60-megadalton plasmid of Escherichia coli serotype O157:H7.
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Enzyme-linked immunosorbent assay for products of the 60-megadalton plasmid of Escherichia coli serotype O157:H7.

机译：大肠杆菌O157：H7血清型60兆尔顿质粒产物的酶联免疫吸附测定。

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Eighty strains of pathogenic Escherichia coli, representing each of the major diarrheal disease-causing groups, were examined by direct enzyme-linked immunosorbent assay (ELISA) for the presence of proteins associated with a 60-MDa plasmid from E. coli serotype O157:H7. Antiserum specific for plasmid-encoded proteins was prepared by immunizing a rabbit with a wild-type E. coli O157:H7 strain (strain 7785) and absorbing the serum with a plasmid-cured derivative (strain 2-45). Use of this antiserum in Western immunoblot analysis detected two proteins of 82 and 92 kDa in strain 7785 but not in strain 2-45. All 16 wild-type E. coli O157:H7 strains and all 10 Shiga-like toxin (SLT)-producing E. coli strains of serotypes other than O157 were ELISA positive. Thirteen of 14 enterotoxigenic and all of 24 enteroinvasive E. coli strains were ELISA negative, as were all of 16 E. coli strains isolated from healthy persons. Of 16 traditional enteropathogenic E. coli (EPEC) serotypes, 10 were ELISA positive, including 10 of 12 strains carrying the EPEC adherence factor gene. Absorption of the serum with an EPEC adherence factor-positive EPEC eliminated EPEC reactivity. This study demonstrates that two plasmid-mediated proteins are common to E. coli O157:H7 strains and to SLT-producing strains of other serotypes. Detection of these proteins by ELISA provides a sensitive and specific screening test for identifying SLT-producing E. coli of both O157 and non-O157 serotypes. Identification of the cross-reactive proteins found in EPEC could provide the basis for a single assay to detect both EPEC and SLT-producing E. coli.
机译：通过直接酶联免疫吸附测定（ELISA）检查了代表每个主要腹泻疾病组的80株致病性大肠杆菌菌株中是否存在与大肠杆菌O157：H7血清型60-MDa质粒相关的蛋白质。通过用野生型大肠杆菌O157：H7菌株（菌株7785）免疫兔子，并用质粒固化的衍生物（菌株2-45）吸收血清来制备对质粒编码蛋白具有特异性的抗血清。在Western免疫印迹分析中使用此抗血清可在7785株中检测到82和92 kDa的两种蛋白质，而在2-45株中未检测到。除O157以外的所有16种血清型野生型大肠杆菌O157：H7菌株和所有10种产生志贺样毒素（SLT）的大肠杆菌菌株均为ELISA阳性。 14株产肠毒素的大肠杆菌中有13株以及24株肠侵害性大肠杆菌菌株均为ELISA阴性，从健康人群中分离出的16株大肠杆菌也都呈ELISA阴性。在16种传统的肠致病性大肠杆菌（EPEC）血清型中，有10种呈ELISA阳性，包括12种带有EPEC粘附因子基因的菌株中的10种。用EPEC粘附因子阳性的EPEC吸收血清可消除EPEC反应性。这项研究表明，两种质粒介导的蛋白是大肠杆菌O157：H7菌株和其他血清型产生SLT的菌株共有的。通过ELISA对这些蛋白质的检测提供了灵敏而特异性的筛选测试，用于鉴定O157和非O157血清型的产生SLT的大肠杆菌。 EPEC中发现的交叉反应蛋白的鉴定可以为检测EPEC和产生SLT的大肠杆菌进行单一测定提供基础。

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