Kim Hyun-Joong, Ryu Ji-Oh, Song Ji-Yeon, Kim Hae-Yeong
Department of Food Science and Biotechnology, Kyung Hee University , Yongin, Republic of Korea.
Foodborne Pathog Dis. 2017 Jul;14(7):400-406. doi: 10.1089/fpd.2016.2221. Epub 2017 Apr 12.
In the detection of Shigella species using molecular biological methods, previously known genetic markers for Shigella species were not sufficient to discriminate between Shigella species and diarrheagenic Escherichia coli. The purposes of this study were to screen for genetic markers of the Shigella genus and four Shigella species through comparative genomics and develop a multiplex polymerase chain reaction (PCR) for the detection of shigellae and Shigella species. A total of seven genomic DNA sequences from Shigella species were subjected to comparative genomics for the screening of genetic markers of shigellae and each Shigella species. The primer sets were designed from the screened genetic markers and evaluated using PCR with genomic DNAs from Shigella and other bacterial strains in Enterobacteriaceae. A novel Shigella quintuplex PCR, designed for the detection of Shigella genus, S. dysenteriae, S. boydii, S. flexneri, and S. sonnei, was developed from the evaluated primer sets, and its performance was demonstrated with specifically amplified results from each Shigella species. This Shigella multiplex PCR is the first to be reported with novel genetic markers developed through comparative genomics and may be a useful tool for the accurate detection of the Shigella genus and species from closely related bacteria in clinical microbiology and food safety.
在使用分子生物学方法检测志贺氏菌属时，先前已知的志贺氏菌属遗传标记不足以区分志贺氏菌属与致泻性大肠杆菌。本研究的目的是通过比较基因组学筛选志贺氏菌属及四种志贺氏菌的遗传标记，并开发一种多重聚合酶链反应（PCR）用于检测志贺氏菌属及志贺氏菌种。对来自志贺氏菌属的总共七个基因组DNA序列进行比较基因组学分析，以筛选志贺氏菌属及每种志贺氏菌的遗传标记。根据筛选出 的遗传标记设计引物组，并使用来自志贺氏菌和肠杆菌科其他细菌菌株的基因组DNA通过PCR进行评估。从评估后的引物组开发出一种新型的志贺氏菌五重PCR，用于检测志贺氏菌属、痢疾志贺氏菌、鲍氏志贺氏菌、福氏志贺氏菌和宋内志贺氏菌，并且通过每种志贺氏菌的特异性扩增结果证明了其性能。这种志贺氏菌多重PCR是首个报道的利用通过比较基因组学开发的新型遗传标记的方法，可能是临床微生物学和食品安全领域中从密切相关细菌中准确检测志贺氏菌属及菌种的有用工具。
