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利用编码β-葡萄糖醛酸酶的基因片段对大肠杆菌和志贺氏菌属进行特异性检测。

Specific detection of Escherichia coli and Shigella species using fragments of genes coding for beta-glucuronidase.

作者信息

Cleuziat P, Robert-Baudouy J

机构信息

Laboratoire de Génétique Moléculaire des Microorganismes, Institut National des Sciences Appliquées, Villeurbanne, France.

出版信息

FEMS Microbiol Lett. 1990 Nov;60(3):315-22. doi: 10.1016/0378-1097(90)90324-j.

Abstract

The occurrence of beta-glucuronidase activity, a main characteristic of Escherichia coli and the presence of the uid chromosomal region of E. coli, coding for this enzyme, were tested on representative members of enteric bacteria. DNA hybridization techniques using uid probes and amplification experiments of uidA gene by the polymerase chain reaction (PCR) confirmed the specificity of uid genes for E. coli and Shigella spp. (i.e., S. boydii, S. dysenteriae, S. flexneri and S. sonnei), independent of the beta-glucuronidase phenotype of bacterial strains. This specificity seemed to be conserved when studies were extended to a wide range of bacteria. It was not possible to distinguish E. coli from Shigella spp. The detection sensitivity using double stranded DNA radiolabeled probes was 3 x 10(4) bacteria and could be brought down to 8 bacteria by PCR. Thus, the uid genes appeared to be ideal candidates for DNA probes technology to detect E. coli-Shigella species.

摘要

对肠道细菌的代表性菌株检测了β-葡萄糖醛酸酶活性的出现情况(这是大肠杆菌的一个主要特征)以及编码该酶的大肠杆菌uid染色体区域的存在情况。使用uid探针的DNA杂交技术以及通过聚合酶链反应(PCR)对uidA基因进行的扩增实验证实了uid基因对大肠杆菌和志贺氏菌属(即鲍氏志贺菌、痢疾志贺菌、福氏志贺菌和宋内志贺菌)的特异性,这与细菌菌株的β-葡萄糖醛酸酶表型无关。当研究扩展到广泛的细菌时,这种特异性似乎是保守的。无法区分大肠杆菌和志贺氏菌属。使用双链DNA放射性标记探针的检测灵敏度为3×10⁴个细菌,通过PCR可降至8个细菌。因此,uid基因似乎是用于检测大肠杆菌-志贺氏菌属的DNA探针技术的理想候选基因。

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