Evaluation of a QIAamp DNA stool purification kit for Shiga-toxigenic Escherichia coli detection in bovine fecal swabs by PCR.
A commercial kit intended for Taq polymerase inhibitor removal was tested to detect Shiga-toxigenic Escherichia coli (STEC) by polymersase chain reaction (PCR) directly from cattle fecal samples. Forty-five samples were analysed for the presence of stx genes. Results were compared to those obtained by two other methods: amplification of DNA purified by a non-commercial procedure (heat lysis protocol), and amplification of DNA from samples cultured in solid media, commonly used in our lab. Identical numbers of positive samples (33/45, 73 %) were obtained with the QIAamp DNA stool purification kit and the culturing procedure, suggesting an adequate removal of inhibitors that interfere in PCR amplification from the feces. Besides, the number of positive samples detected using DNA purified by the non-commercial protocol was lower, 25/39 (64%) than that achieved by using the kit. In conclusion, the use of the QIAamp DNA stool purification kit provided a rapid stx gene detection by PCR in bovine fecal samples.
Gioffré
A
A
Institute of Biotechnology, CNIA-INTA, 1712 Castelar, Argentina.
Meichtri
L
L
Zumárraga
M
M
Rodríguez
R
R
Cataldi
A
A
D000818:Animals; D002417:Cattle / Q000382:microbiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000302:isolation & purification* / Q000378:metabolism; D005243:Feces / Q000382:microbiology*; D005506:Food Contamination / Q000517:prevention & control; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D012007:Rectum / Q000382:microbiology; D012680:Sensitivity and Specificity; D022603:Shiga Toxins / Q000032:analysis*; D019914:Taq Polymerase / Q000037:antagonists & inhibitors
D004791:Enzyme Inhibitors; D011933:Reagent Kits, Diagnostic; D022603:Shiga Toxins; D019914:Taq Polymerase