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Ancylostoma ceylanicum ,  A. caninum , and  Giardia lamblia  assemblage A are common intestinal parasites of dogs and cats; they can also infect humans, causing parasitic zoonoses. In this study, a multiplex PCR method was developed for simultaneous identification and detection of those three zoonotic parasites. Three pairs of specific primers were designed based on ITS sequence of  A. ceylanicum  and  A. caninum  and TPI gene of  G. lamblia  available in the GenBank. The multiplex PCR reaction system was established by optimizing the reaction condition, and a series of tests on the sensitivity, specificity, and clinical application were also conducted. Results showed that three target fragments were amplified specifically; the detection limit was 10 eggs for both  A. ceylanicum  and  A. caninum , 72 pg DNA for  G. lamblia . Of 112 clinical fecal samples, 34.8% and 17.8% samples were positive for  A. caninum  and  A. ceylanicum , respectively, while only 2.7% samples were positive for  G. lamblia  assemblage A. It is concluded that the established multiplex PCR assay is a convenient, rapid, cost-effective, and high-efficiency method for molecular detection and epidemiological investigation of three zoonotic parasites.

biomed research international

A Multiplex PCR for Simultaneous Detection of Three Zoonotic ParasitesAncylostoma ceylanicum,A. caninum,andGiardia lambliaAssemblage A