Non-radioactive oligonucleotide probe for detection of clinical enterotoxigenic Escherichia coli isolates of bovine origin.
Oligonucleotides (32 or 34 mer) corresponding to enterotoxigenic Escherichia coli STIa (ST-P) toxin gene were tailed with Bio-11-dUTP using terminal deoxynucleotidyl transferase. Plasmids from clinical 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 probes 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.
D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics; D001710:Biotin; D002417:Cattle / Q000382:microbiology; D004768:Enterotoxins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D004927:Escherichia coli Infections / Q000175:diagnosis*; D029968:Escherichia coli Proteins; D015345:Oligonucleotide Probes
D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D015345:Oligonucleotide Probes; C034151:heat stable toxin (E coli); D001710:Biotin