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用合成寡核苷酸探针检测大肠杆菌肠毒素编码基因中的核苷酸序列变异。

Detection with synthetic oligonucleotide probes of nucleotide sequence variations in the genes encoding enterotoxins of Escherichia coli.

作者信息

Nishibuchi M, Murakami A, Arita M, Jikuya H, Takano J, Honda T, Miwatani T

机构信息

Research Institute for Microbial Diseases, Osaka University, Japan.

出版信息

J Clin Microbiol. 1989 Oct;27(10):2272-6. doi: 10.1128/jcm.27.10.2272-2276.1989.

Abstract

We examined variations in the genes encoding heat-stable enterotoxin (ST) and heat-labile enterotoxin (LT) in 88 strains of Escherichia coli isolated from individuals with traveler's diarrhea to find suitable sequences for use as oligonucleotide probes. Four oligonucleotide probes of the gene encoding ST of human origin (STIb or STh), one oligonucleotide probe of the gene encoding ST of porcine origin (STIa or STp), and three oligonucleotide probes of the gene encoding LT of human origin (LTIh) were used in DNA colony hybridization tests. In 15 of 22 strains possessing the STh gene and 28 of 42 strains producing LT, the sequences of all regions tested were identical to the published sequences. One region in the STh gene examined with a 18-mer probe was relatively well conserved and was shown to be closely associated with the enterotoxicity of the E. coli strains in suckling mice. This oligonucleotide, however, hybridized with strains of Vibrio cholerae O1, V. parahaemolyticus, and Yersinia enterocolitica that gave negative results in the suckling mouse assay.

摘要

我们检测了从旅行者腹泻患者中分离出的88株大肠杆菌中编码热稳定肠毒素(ST)和热不稳定肠毒素(LT)的基因变异情况,以寻找适合用作寡核苷酸探针的序列。在DNA菌落杂交试验中,使用了四种源自人类的ST编码基因(STIb或STh)的寡核苷酸探针、一种源自猪的ST编码基因(STIa或STp)的寡核苷酸探针以及三种源自人类的LT编码基因(LTIh)的寡核苷酸探针。在拥有STh基因的22株菌株中的15株以及产生LT的42株菌株中的28株中,所有检测区域的序列均与已发表序列相同。用18聚体探针检测的STh基因中的一个区域相对保守,并且显示与哺乳小鼠中大肠杆菌菌株的肠毒性密切相关。然而,该寡核苷酸与在哺乳小鼠试验中呈阴性结果的霍乱弧菌O1、副溶血性弧菌和小肠结肠炎耶尔森菌菌株发生杂交。

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