Use of a multiplex PCR system for the simultaneous detection of heat labile toxin I and heat stable toxin II genes of enterotoxigenic Escherichia coil in skim milk and porcine stool.
Enterotoxigenic Escherichia coli (ETEC) strains which produce heat labile and/or heat stable toxins (LT and ST) may cause diarrhea in humans and farm animals. Using PCR primers specific for the LT I and ST II genes, a multiplex PCR system which allows detection of LT 1-and ST 11-producing ETEC strains was developed. When skim milk was used for a PCR assay, it was found that if target cells in the sample were precultured in MacConkey broth for 8 h prior to PCR as few as 10(0) cells per ml of the sample could be detected. Without the preculture step, 10(4) CFU of target cells per 0.2 g of porcine stool specimen were required to generate visible PCR products. The multiplex PCR system can be used for rapid testing of fecal specimens, food and possibly environmental samples for the presence of ETEC strains.
Department of Food Science, National Chung-Hsing University Taichung, Taiwan, ROC
H Y
HY
Tsen
L Z
LZ
Jian
W R
WR
Chi
D000818:Animals; D001427:Bacterial Toxins / Q000032:analysis / Q000235:genetics*; D015169:Colony Count, Microbial; D017931:DNA Primers; D004768:Enterotoxins / Q000032:analysis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000472:pathogenicity; D029968:Escherichia coli Proteins; D005243:Feces / Q000382:microbiology*; D008892:Milk / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013552:Swine / Q000382:microbiology*
D001427:Bacterial Toxins; D017931:DNA Primers; D004768:Enterotoxins; D029968:Escherichia coli Proteins; C034151:heat stable toxin (E coli); C027538:heat-labile enterotoxin, E coli