建立一种用于区分大肠杆菌和志贺氏菌属的双重实时 PCR 方法
Development of a duplex real-time PCR for differentiation between E. coli and Shigella spp.
机构信息
Bavarian Health and Food Safety Authority, Oberschleissheim, Germany.
出版信息
J Appl Microbiol. 2011 May;110(5):1245-51. doi: 10.1111/j.1365-2672.2011.04973.x. Epub 2011 Mar 14.
AIMS
The aim of this study was to develop a real-time PCR test for differentiation between Shigella spp. and E. coli, in particular enteroinvasive Escherichia coli (EIEC).
METHODS AND RESULTS
A duplex real-time PCR specific for the genes encoding for β-glucuronidase (uidA) and lactose permease (lacY) was developed. Ninety-six isolates including 11 EIEC isolates of different serotypes and at least three representatives of each Shigella species were used for selectivity testing. All isolates tested were positive for the uidA gene. Additionally, all E. coli isolates were positive for the lacY gene, whereas no Shigella isolate tested harboured lacY.
CONCLUSIONS
The duplex real-time PCR assay was found to be simple, rapid, reliable and specific.
SIGNIFICANCE AND IMPACT OF THE STUDY
If possible at all, delineation of so-called inactive EIEC from Shigella spp. is cumbersome. Biochemical and serological methods are limited to specific pheno- and serotypes. This assay clearly simplifies the differentiation of both.
目的
本研究旨在开发一种实时 PCR 检测方法，用于区分志贺氏菌属和大肠杆菌，特别是侵袭性大肠杆菌（EIEC）。
方法和结果
开发了一种针对编码β-葡萄糖醛酸酶（uidA）和乳糖通透酶（lacY）基因的双重实时 PCR 方法。使用了 96 株分离株，包括 11 种不同血清型的 EIEC 分离株和每种志贺氏菌属的至少 3 个代表进行选择性测试。所有测试的分离株 uidA 基因均呈阳性。此外，所有大肠杆菌分离株 lacY 基因均呈阳性，而没有志贺氏菌分离株携带 lacY。
结论
该双重实时 PCR 检测方法简单、快速、可靠且具有特异性。
研究的意义和影响
如果可能的话，将所谓的不活跃的 EIEC 与志贺氏菌属区分开来是很麻烦的。生化和血清学方法仅限于特定的表型和血清型。该检测方法明显简化了两者的区分。
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