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通过DNA菌落杂交法检测产肠毒素大肠杆菌

Detection of enterotoxigenic Escherichia coli by DNA colony hybridization.

作者信息

Moseley S L, Huq I, Alim A R, So M, Samadpour-Motalebi M, Falkow S

出版信息

J Infect Dis. 1980 Dec;142(6):892-8. doi: 10.1093/infdis/142.6.892.

Abstract

A method fo detecting large numbers of isolates of enterotoxigenic Escherichia coli is described in which the genes encoding th enterotoxins are detected, rather than the toxins themselves. Radiolabeled fragments of DNA encoding the heat-labile (LT) or heat-stable (ST) toxins were used as hybridization probes for homologous DNA sequences in E. coli colonies grown and lysed in situ on nitrocellulose filters. The LT probe detected all of 31 E. coli strains producing ST and LT or only LT, while the ST probe detected 12 of 17 strains producing only ST and three of 26 strains producing ST and LT. These results suggest that the LTs produced by different isolates of E. coli are homologous and that human isolates of E. coli produce at least two heterologous STs detectable in the infant mouse assay. The hybridization method also detected the presence of enterotoxigenic E. coli in bacterial growth in directly spotted stools from patients with acute diarrhea.

摘要

本文描述了一种检测大量产肠毒素大肠杆菌分离株的方法,该方法检测的是编码肠毒素的基因,而非毒素本身。编码不耐热(LT)或耐热(ST)毒素的放射性标记DNA片段被用作杂交探针,用于检测在硝酸纤维素滤膜上原位生长并裂解的大肠杆菌菌落中的同源DNA序列。LT探针检测出了31株产生ST和LT或仅产生LT的大肠杆菌菌株,而ST探针检测出了17株仅产生ST的菌株中的12株以及26株产生ST和LT的菌株中的3株。这些结果表明,不同大肠杆菌分离株产生的LT是同源的,并且人类大肠杆菌分离株产生至少两种在幼鼠试验中可检测到的异源ST。该杂交方法还检测出急性腹泻患者直接点样粪便中的细菌生长物中存在产肠毒素大肠杆菌。

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