Detection of enterotoxigenic Escherichia coli in stool specimens by polymerase chain reaction.
A polymerase chain reaction (PCR) protocol for rapid (7 h) detection of enterotoxigenic Escherichia coli (ETEC) is described. This protocol has been validated on 57 stool samples from young children by comparing it with the colony hybridization technique. A good agreement was found between the two methods with Cohen's kappa statistics of 0.87 and 0.79 for the detection of the heat-stable toxin (ST) and heat-labile toxin (LT), respectively. Of 26 samples positive for LT and 15 samples positive for ST by colony hybridization, 21 (81%) and 15 (100%) were also found to be positive for LT and ST by PCR, respectively. Only one sample identified as LT-negative by colony hybridization was found to be positive by PCR. However, 3 of 42 samples of ST-negative by colony hybridization were detected as positive by PCR. A reconstruction experiment revealed that PCR could detect LT-producing and ST-producing ETEC at minimal concentrations of 2.5 x 10(3) cfu and 2.5 x 10(2) cfu per gram of feces, respectively. These data indicate the possible use of this method for rapid identification of ETEC-associated diarrhea in clinical and epidemiological settings.
Yavzori
M
M
Medical Corps, Israel Defence Force, Medical School, Ben-Gurion University, Israel.
Porath
N
N
Ochana
O
O
Dagan
R
R
Orni-Wasserlauf
R
R
Cohen
D
D
D001427:Bacterial Toxins / Q000096:biosynthesis / Q000235:genetics*; D002648:Child; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D003967:Diarrhea / Q000382:microbiology*; D004768:Enterotoxins / Q000096:biosynthesis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D004927:Escherichia coli Infections / Q000382:microbiology; D029968:Escherichia coli Proteins; D005243:Feces / Q000382:microbiology*; D006801:Humans; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D001427:Bacterial Toxins; D015342:DNA Probes; D004269:DNA, Bacterial; D004768:Enterotoxins; D029968:Escherichia coli Proteins; C034151:heat stable toxin (E coli); C027538:heat-labile enterotoxin, E coli