Abstract: The primers and probes were designed and synthesized according to the sequences of transmissible gastroenteritis virus and β-actin, and then reaction parameters were optimized to develop a TaqMan fluorescence quantitative RT-PCR assay. Meanwhile,37 field samples were detected and the results were compared with that of routine RT-PCR and antigen rapid test kit. It was showed that the fluorescence quantitative RT-PCR assay could detect 15.3copies/µL of plasmid DNA and its specificity and reproducibility were very good，while the sensitivity of the routine RT-PCR was 1.53×103copies/µL. The result of field test also showed that its sensitivity was higher than that of the routine RT-PCR and TGEV antigen rapid test kit (chromatographic immunoassay).