Polymerase chain reaction for detection of verocytotoxigenic Escherichia coli isolated from animal and food sources.
Animals and their by-products have been implicated as important sources of verocytotoxigenic Escherichia coli (VTEC) associated with disease in humans. VTEC comprise a wide range of serotypes and produce a variety of closely related verocytotoxins (VT). A pair of oligonucleotide primers, targeting conserved sequences found in VT1, VT2 and VTE genes, was used to develop a polymerase chain reaction (PCR) procedure to detect all types of VTEC. Supernatants of boiled broth cultures of VTEC (223 strains) isolated from ground beef, ground pork, raw milk, bovine faeces and porcine faeces; non-VTEC E. coli (72 strains); and other enteric and food bacteria (76 strains) were tested by PCR. The verocytotoxigenicity of these strains was verified by the Vero cell assay. All 223 VTEC isolates, comprising over 50 different serotypes, were detected by the PCR procedure. Shigella dysenteriae type 1 was the only other bacterium that was positive in this assay. As little as 1 pg of VTEC DNA and as few as 17 cfu of VTEC could be detected with this method. The results indicate that these primers detect VTEC over a wide range of serotypes. This method may be applicable as a screening procedure for the detection of VTEC in samples of foods and faeces.
Read
S C
SC
Agriculture Canada, Health of Animals Laboratory, Guelph, Ontario.
Clarke
R C
RC
Martin
A
A
De Grandis
S A
SA
Hii
J
J
McEwen
S
S
Gyles
C L
CL
D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics*; D002417:Cattle / Q000382:microbiology; D003603:Cytotoxins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D005516:Food Microbiology; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2; D013552:Swine / Q000382:microbiology; D014709:Vero Cells
D001427:Bacterial Toxins; D003603:Cytotoxins; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2