1887

Abstract

Differentiation between and is problematic in clinical specimens due to fastidious growth requirements and limited biochemical tests. This study describes a rapid, multiplex PCR protocol for the direct detection and differentiation of and in stools. An evaluation was carried out of this multiplex protocol based on the detection of (genus specific), and () and () genes, using stool from patients with enteritis and chicken. Protocol sensitivity was assessed and specificity determined using a panel of enteric bacteria, and evaluation of 30 diarrhoeic stool specimens culture negative for . Of the 114 specimens (54 human and 60 chicken) evaluated by the protocol, 70 (61.4 %) were identified as , 35 (30.7 %) as and 9 (7.9 %) as a mixed infection/colonization with both species. All mixed infections were identified as by culture. Among the stool specimens that were culture negative for , two (6.7 %) were positive by multiplex PCR. The protocol sensitivity limit was 0.015–0.016 ng and DNA μl in the specimen. There was no cross-reaction with the reference strains assessed. Comparison of hippurate test and multiplex PCR demonstrated 17 isolates with false-positive hippurate enzymic activity and 7 with false-negative activity. This rapid protocol (turnaround time 6 h) is highly sensitive and specific for direct evaluation of stool for these pathogens. It has significant application for routine clinical diagnostic and epidemiological purposes.

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2007-10-01
2019-11-22
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