用于鉴定肠出血性大肠杆菌的酶标寡核苷酸探针的评估

Evaluation of enzyme-labeled oligonucleotide probes to identify enterohaemorrhagic Escherichia coli.

作者信息

Yoh M, Takagi K, Eda J, Ohtomo M, Takarada Y, Shibata S, Honda T

机构信息

Research Center for Emerging Infectious Diseases, Osaka University, Japan. myon@bikin, osaka-u.ac.jp

出版信息

Microbiol Immunol. 1997;41(11):879-82. doi: 10.1111/j.1348-0421.1997.tb01944.x.

DOI:10.1111/j.1348-0421.1997.tb01944.x

PMID:9444330

Abstract

Alkaline phosphatase-conjugated oligonucleotide probes were developed to detect the gene coding for Vero toxin 1 (VT1) and Vero toxin 2 (VT2). Using these probes, 3 hr was enough to detect VT genes when suspicious colonies of enterohaemorrhagic Escherichia coli (EHEC) were obtained on an agar plate. The results of a hybridization test with 144 isolates of EHEC O157 and one isolate of Shigella dysenteriae Type 1 agreed exactly with the immunological detection, reversed passive latex agglutination (RPLA) test, of VTs in their culture supernatants. The sensitivity levels of these probes for the detection of VT genes were 100%. The specificity of these probes were also tested with a total of 1,002 strains of Escherichia coli other than EHEC and 8 strains of Shigella sp. other than Shigella dysenteriae Type 1; the results showed 100% specificity.

摘要

开发了碱性磷酸酶偶联的寡核苷酸探针来检测编码维罗毒素1（VT1）和维罗毒素2（VT2）的基因。使用这些探针，当在琼脂平板上获得肠出血性大肠杆菌（EHEC）的可疑菌落时，3小时就足以检测VT基因。对144株EHEC O157分离株和1株痢疾志贺菌1型分离株进行杂交试验的结果与它们培养上清液中VTs的免疫检测——反向被动乳胶凝集（RPLA）试验完全一致。这些探针检测VT基因的灵敏度水平为100%。还使用总共1002株非EHEC的大肠杆菌菌株和8株非痢疾志贺菌1型的志贺菌属菌株测试了这些探针的特异性；结果显示特异性为100%。