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使用非同位素和同位素标记通过菌落杂交法评估沙门氏菌特异性DNA探针。

Evaluation of a Salmonella-specific DNA probe by colony hybridization using non-isotopic and isotopic labeling.

作者信息

Aabo S, Thomas A, Hall M L, Smith H R, Olsen J E

机构信息

Department of Veterinary Microbiology, Royal Veterinary and Agricultural University, Denmark.

出版信息

APMIS. 1992 Jul;100(7):623-8. doi: 10.1111/j.1699-0463.1992.tb03976.x.

Abstract

A 2.3 kilobase (kb) Salmonella probe, JEO402-1, and two subfragments, F1214 (1.3 kb) and F1217 (0.8 kb), have been evaluated by colony hybridization using pure cultures of Salmonella serovars and non-salmonella bacteria. JEO402-1, and its subfragments, F1214 and F1217, hybridized to all of 156 different Salmonella serovars tested, while there was no reaction to 112 non-salmonella strains belonging to 19 genera and 37 species of Enterobacteriaceae. Together with previously published results, the JEO402-1 probe has now been shown to detect a total of 396 Salmonella strains belonging to 214 serovars of Salmonella subspecies I-VI. A total of 178 non-salmonella strains representing 23 genera and 51 species of Enterobacteriaceae have all tested negative with JEO402-1. The hybridization results obtained using a digoxigenin-labeled probe were similar to those obtained with 35S isotopic labeling when complete colony lysis was ensured.

摘要

一个2.3千碱基(kb)的沙门氏菌探针JEO402 - 1以及两个亚片段F1214(1.3 kb)和F1217(0.8 kb),已通过使用沙门氏菌血清型纯培养物和非沙门氏菌进行菌落杂交评估。JEO402 - 1及其亚片段F1214和F1217与所测试的156种不同沙门氏菌血清型全部杂交,而对属于肠杆菌科19个属和37个种的112株非沙门氏菌菌株无反应。连同先前发表的结果,现在已表明JEO402 - 1探针可检测总共属于沙门氏菌亚种I - VI的214个血清型的396株沙门氏菌菌株。总共代表肠杆菌科23个属和51个种的178株非沙门氏菌菌株用JEO402 - 1检测均为阴性。当确保完全菌落裂解时,使用地高辛标记探针获得的杂交结果与用35S同位素标记获得的结果相似。

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