Abstract: Objective Comparative the results of PCR and serology methods for detecting of three different pathogenes. Methods The nucleic acid of Toxoplasma gondii、Brucella and Rotavirus extracted from the liver of ICR mice、stool speciments and vagina specimen of Beagle、stool specimen of Macaca mulata respectively was amplified by NT-PCR、PCR and RT-PCR. The serum antibody of Toxoplasma gondii、Brucella and Rotavirus from ICR mice、Beagle and Macaca mulata respectively was detected by serology methods. Results The results of NT-PCR、PCR、RT-PCR were accorded with serology methods.Conclusion The developed NT-PCR、PCR、RT-PCR assay was a rapid, sensitive and specific method, it can be used in monitoring pathogen.