Non-radioactive oligonucleotide probe for detection of clinical enterotoxigenic Escherichia Coli isolates of bovine origin |
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| Affiliation: | 1. Department of Building, Civil and Environmental Engineering, Concordia University, Montreal, QC H3G1M8, Canada;2. Department of Civil and Environmental Engineering, University of California Los Angeles, Los Angeles, CA 90095-1593, United States;1. Interuniversity Department of Regional and Urban Studies and Planning, Politecnico di Torino, Viale Mattioli, 39, Torino 10125, Italy;1. School of Mechanical Engineering, Xi’an Jiaotong University, Xi’an 710049, PR China;2. State Key Laboratory for Manufacturing Systems Engineering, Xi’an Jiaotong University, Xi’an 710054, PR China |
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| Abstract: | ![]()
Oligonucleotides (32 or 34 mer) corresponding to enterotoxigenic Escherichia coli STIa (ST-P) toxin were tailed with Bio-11-dUTP using terminal deoxynucleotidyl transferase. Plasmids from clinica E.Coli isolates were prepared by modified rapid alkaline lysis procedure and dot-spotted. Biotinylated oligonucleotide probes were hybridized to detect the StIa toxin gene. The results were in agreement with that obtained by radioactive oligonucleotide probes. Of 135 clinical isolates (sampled from 6 different regions of France), only 7 (5.2 %) were found to be STIa+. These 7 isolates were the only ones to be found positive for the K99 adhesive pili antigen. Both the proves were specific to the STIa toxin gene and failed to detect the closely related STIb (ST-H) toxin gene. Possibilities of their wide usage in clinical labs are discussed. |
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