Establishment of a TaqMan multiplex quantitative PCR method for detecting three different serovars of Glaesserella parasuis serovars 5/12, 7 and 11.

Abstract

Glaesserella parasuis, a commensal in the upper respiratory tract of swine, can cause Glässer's disease in the stress conditions, which caused great economic loss to the swine industry. So far, 15 different serovars have been identified, and which are unevenly distributed in different countries and regions. To date, dominant serovars of 5/12, 7, and 11 have significantly increased in China. To rapidly and simultaneously detect these 3 dominant serovars, a TaqMan multiplex quantitative PCR (qPCR) detection method was developed. The study designed primers and probes based on the conserved sequences of the specific genes wcwK, funQ, and amtA for serovars 5/12, 7, and 11, respectively, the reaction and conditions for single and triplex systems were respectively optimized. A TaqMan multiplex qPCR method was successfully established that can simultaneously identify these three serovars. The specificity and sensitivity of the results indicated that the detection limit for the concentrations of the wcwK, funQ, and amtA genes was 10copies/µL, and there was no cross-reaction with other common pathogens. The successful establishment of this qPCR method will provide a convenient and rapid method for the epidemic status investigation of serovars 5/12, 7 and 11.