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用于菌落杂交的克隆多核苷酸和合成寡核苷酸探针在鉴定产肠毒素大肠杆菌方面同样有效。

Cloned polynucleotide and synthetic oligonucleotide probes used in colony hybridization are equally efficient in the identification of enterotoxigenic Escherichia coli.

作者信息

Sommerfelt H, Kalland K H, Raj P, Moseley S L, Bhan M K, Bjorvatn B

机构信息

Institute of International Health, University of Bergen, Norway.

出版信息

J Clin Microbiol. 1988 Nov;26(11):2275-8. doi: 10.1128/jcm.26.11.2275-2278.1988.

Abstract

Restriction endonuclease-generated polynucleotide and synthetically produced oligonucleotide gene probes used in colony hybridization assays proved to be efficient for the detection and differentiation of enterotoxigenic Escherichia coli. To compare their relative efficiencies, these two sets of probes were radiolabeled with 32P and were applied to 74 strains of E. coli with known enterotoxin profiles and to 156 previously unexamined E. coli isolates. The enterotoxigenic bacteria Vibrio cholerae O1, Vibrio cholerae non-O1 (NAG), Yersinia enterocolitica, and E. coli harboring the plasmid vectors of the polynucleotide gene probes were examined for further evaluation of probe specificity. The two classes of probes showed a perfect concordance in their specific detection and differentiation of enterotoxigenic E. coli. In the analysis of six strains, the signal strength on autoradiography after hybridization with oligonucleotides was weaker than that obtained after hybridization with polynucleotide probes. The probes did not hybridize with DNA from V. cholerae O1, V. cholerae non-O1 (NAG), or Y. enterocolitica. The strains of E. coli harboring the plasmid vectors of the polynucleotide gene probes were, likewise, negative in the hybridization assays.

摘要

在菌落杂交试验中使用的限制性内切酶产生的多核苷酸和化学合成的寡核苷酸基因探针,被证明对产肠毒素大肠杆菌的检测和鉴别是有效的。为了比较它们的相对效率,这两组探针用³²P进行放射性标记,并应用于74株已知产肠毒素谱的大肠杆菌菌株以及156株先前未经检测的大肠杆菌分离株。对产肠毒素细菌霍乱弧菌O1、非O1霍乱弧菌(NAG)、小肠结肠炎耶尔森菌以及携带多核苷酸基因探针质粒载体的大肠杆菌进行检测,以进一步评估探针的特异性。这两类探针在产肠毒素大肠杆菌的特异性检测和鉴别方面表现出完全一致的结果。在对6株菌株的分析中,与寡核苷酸杂交后放射自显影的信号强度弱于与多核苷酸探针杂交后获得的信号强度。这些探针不与霍乱弧菌O1、非O1霍乱弧菌(NAG)或小肠结肠炎耶尔森菌的DNA杂交。同样,携带多核苷酸基因探针质粒载体的大肠杆菌菌株在杂交试验中也呈阴性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/02e8/266875/4f7553528cb7/jcm00083-0059-a.jpg

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