Establishment of Taqman real-time PCR for rapid detection of Wohlfahrtiimonas chitiniclastica bacilli

Objective To develop a Taqman real-time polymerase chain reaction(PCR) method for the determination of Wohlfahrtiimonas chitiniclastica(W.chitiniclastica).Methods Primers and specific fluorogenic probes(TaqMan) were designed for W.chitiniclastica with the Primer Express software based on the genome sequence of W.chitiniclastica SH04 strain; then the specificity, sensitivity, and repeatability of the method were verified.Results The method has goog specificity, with the coefficient of variation(CV) values of less than 2%in repetitive testing and a sensitivity value of 19 colony forming unit(CFU) in a 20μL reaction system.Using the method established, the detection results for 6 artificially W.chitiniclastica-polluted samples were all positive.Conclusion The real-time PCR method is a specific, sensitive and reproducible assay for the detection of W.chitiniclastica.