Objective To establish a real-time fluorescent quantitative PCR (Q-PCR) method for detection of feline herpesvirus 1(FHV-1)in experiment cats and clinical sick cats. Methods Primers and TaqMan probes were designed and synthesized according to the published FHV-1 specific sequences of TK gene. FHV DNA standards were prepared using molecular biological techniques. The linearity, specificity, sensitivity, stability of the established Q-PCR method were tested. The method was used to detect 48 samples of cats. Results The linear range was 10² copies/μL to 10⁹ copies/μL. The developed Q-PCR method showed no cross reaction with herpes virus type 1 (HSV-1), canine herpesvirus (CHV), pig pseudo rabies virus (PRV) and cat parvovirus (FPV). The sensitivity was 10 copies/μL. The coefficient of variation (CV) was less than 5%. There were 33 positive cases detected in the 48 samples of cats. Conclusions The developed Q-PCR method is good in linearity, specificity, sensitivity, stability, and may be used for rapid quantitative detection of FHV-1 in cats.