Yamamoto K, Honda T, Miwatani T, Tamatsukuri S, Shibata S
Department of Bacteriology and Serology, Osaka University, Japan.
Can J Microbiol. 1992 May;38(5):410-6. doi: 10.1139/m92-069.
Alkaline phosphatase conjugated oligonucleotide probes were developed to detect the genes (tdh and trh) coding for the thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) of Vibrio parahaemolyticus. Using dot blot hybridization, probes were tested with 94 clinical isolates of V. parahaemolyticus. Results agreed well with those obtained using radio-labeled recombinant DNA probes for the genes tdh and trh. Specificity and sensitivity of enzyme tdh probes for detection of the trh gene were 100 and 93%, respectively, and those of the trh probes for trh gene detection were 93 and 86%, respectively. The tdh probes also hybridized with tdh-like genes processed by all strains of V. hollisae, and some strains of V. mimicus and V. cholerae non-O1, but neither tdh nor trh probes reacted with other bacterial species isolated from diarrheal stools. However, some V. parahaemolyticus strains that were negative with the enzyme trh probe hybridized weakly with a radio-labeled trh DNA fragment probe at medium stringency, and a few strains that were negative in high stringency conditions with a radio-labeled trh DNA fragment probe hybridized with the enzyme trh probe. This suggests that some strains of V. parahaemolyticus may carry another gene resembling trh.
开发了碱性磷酸酶偶联的寡核苷酸探针，用于检测副溶血性弧菌耐热直接溶血素（TDH）和TDH相关溶血素（TRH）的编码基因（tdh和trh）。使用斑点印迹杂交法，用94株副溶血性弧菌临床分离株对探针进行了检测。结果与使用放射性标记的重组DNA探针检测tdh和trh基因所获得的结果非常吻合。酶标tdh探针检测trh基因的特异性和敏感性分别为100%和93%，trh探针检测trh基因的特异性和敏感性分别为93%和86%。tdh探针还与所有霍利斯弧菌菌株以及一些模仿弧菌和非O1群霍乱弧菌菌株携带的tdh样基因杂交，但tdh和trh探针均未与从腹泻粪便中分离出的其他细菌物种发生反应。然而，一些用酶标trh探针检测为阴性的副溶血性弧菌菌株，在中等严谨度下与放射性标记的trh DNA片段探针弱杂交，少数在高严谨度条件下与放射性标记的trh DNA片段探针检测为阴性的菌株，与酶标trh探针杂交。这表明一些副溶血性弧菌菌株可能携带另一个类似于trh的基因。
