Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano-shi, Osaka, Japan; Technical Research Laboratory, KURABO INDUSTRIES LTD., Neyagawa, Osaka, Japan.
Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano-shi, Osaka, Japan.
J Microbiol Methods. 2019 Jun;161:111-117. doi: 10.1016/j.mimet.2019.04.016. Epub 2019 May 2.
We developed a new GeneFields® EHEC/SS PCR dipstick DNA chromatography kit for the simultaneously detection of invA, ipaH, and stx genes in Salmonella enterica (56 strains), Shigella spp. (44), and enterohemorrhagic Escherichia coli (EHEC) (28), respectively, and evaluated the sensitivity and specificity with other bacteria (57) by this kit. The sensitivity and specificity were 100%, respectively. The detection limit of various methods was determined using 5% (w/v) stool suspensions spiked with each bacterium. The detection limit of the GeneFields® EHEC/SS kit ranged from approximately 10-10 CFU/g. Additionally, the relative sensitivities and specificities of the GeneFields® EHEC/SS kit vs two commercially available real-time PCR kits were >85.0% and >90.0%, respectively. These results indicate that the GeneFields® EHEC/SS kit can be used for genetic screening of S. enterica, Shigella spp., and EHEC in human stool specimens with sensitivities and specificities similar to those of the commercially available real-time PCR kits.
我们开发了一种新的 GeneFields® EHEC/SS PCR 条带 DNA 色谱试剂盒，用于分别同时检测沙门氏菌属（56 株）、志贺氏菌属（44 株）和肠出血性大肠杆菌（EHEC）（28 株）中的 invA、ipaH 和 stx 基因，并通过该试剂盒评估了与其他细菌（57 株）的灵敏度和特异性。灵敏度和特异性分别为 100%。通过使用含有每种细菌的 5%（w/v）粪便悬浮液来确定各种方法的检测限。GeneFields® EHEC/SS 试剂盒的检测限范围约为 10-10 CFU/g。此外，GeneFields® EHEC/SS 试剂盒与两种市售实时 PCR 试剂盒的相对灵敏度和特异性均>85.0%和>90.0%。这些结果表明，GeneFields® EHEC/SS 试剂盒可用于人粪便标本中沙门氏菌属、志贺氏菌属和 EHEC 的基因筛查，其灵敏度和特异性与市售实时 PCR 试剂盒相似。
