Detection of toxigenic <I>Clostridium difficile</I> in pig feces by PCR
Alvarez-Perez, S. | Alba, P. | Blanco, J.L. | Garcia, M.E.
: <I>Clostridium difficile</I> is considered an important uncontrolled cause of neonatal diarrhea. Also, the presence of bacteria in the feces of the animal could represent a zoonosic risk for the contamination of meat products. Therefore, it is necessary to have procedures available for the early detection of <I>C. difficile</I> in animals. The current study describes a new semi-automated procedure for the recovery of <I>C. difficile</I> DNA from pig feces and subsequent amplification by polymerase chain reaction (PCR) of three different sequences: the triose phosphate isomerase gene <I>tpi</I>, specific for this bacterial species, and the <I>tcdA</I> and <I>tcdB</I> genes, which code for the A and B toxins of <I>C. difficile</I>, respectively. Twenty-two fecal samples microbiologically positive for <I>C. difficile</I> were used. The <I>tpi</I> and <I>tcdA</I> genes were amplified in all of them. The internal fragment of <I>tcdB</I> was detected from 21 of these extracts; the negative sample gave a positive result when a different primer pair was used. None of the 10 DNA extracts obtained from culture-negative samples gave a positive result. The method presented in this article eliminates the interference caused by the possible presence of PCR inhibitors. To the authors’ knowledge, this is the first description of a PCR procedure for detection of <I>C. difficile</I> DNA from domestic animal feces.
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