Abstract: Objective To establish a real-time fluorescent quantitative PCR (Q-PCR) method for detection of mouse hepatitis virus (MHV) in naked mole-rats and other rodent animals. Methods The specific primers and probe were designed and synthesized by selecting conserved regions of MHV E gene to establish the Q-PCR method. Then the linearity, specificity, sensitivity, repeatability and stability of this method was verified. Meanwhile, the established method was used to detect 79 clean grade mice, 63 naked mole-Crats, 35 SPF mice, 10 Mongolian gerbils, 20 golden hamsters and 4 ground squirrels for MHV. Results The real-time fluorescent quantitative PCR method for MHV was successfully established. The linear range of the method was 10¹ copies/μL to 10⁹copies/μL. The method has no cross reaction with Sendai (SV), reovirus type 3 (Reo3), pneumonia virus of mice (PVM), bovine coronavirus (BCV).The detection sensitivity could reach 10 copies/μL, the results of repeatability and stability showed that the coefficient of variation between experiments was less than 5%.The positive rate of MHV in 79 clean grade mice was 22.78%, and that of 63 naked mole-Crats, 35 SPF mice, 10 Mongolian gerbis, 20 golden hamsters and 4 ground squirrels were all negative. Established method was compared with RT-PCR, the coincidence rate was 97.47%. Conclusion The established Q-PCR method for MHV is good in linearity and specificity, it can be used for the detection MHV in multiple varieties rodents.

Key words: Mouse hepatitis virus, Real-time fluorescent quantitative PCR, Rodents