关键词:

小鼠肺炎病毒, RT-PCR, 实验动物, 应用

Abstract:

Objective To establish a RT-PCR method for detection of Pneumonia virus of mice ( PVM ) in laboratory animals and related samples. Method The primers were designed and synthesized by selecting the conserved G gene sequence of PVM, and the RT-PCR method was established. And the specificity, sensitivity,repeatability and stability of the established method were verified. The established method was used to detect the lung tissue samples of 27 mice,9 rats, 5 gerbils submitted for routine examination and 19 PVM infected mice, and 8 ICLAS international comparison samples. Result The established method had no cross reaction with Sendai virus( SV) , Reovirus type 3 ( Reo3) , ( HV) and Lymphocytic Choriomeningitis Virus ( LCMV) ; The method could detect the minimum template concentration of PVM DNA was 8. 77×102 copies / μL, and could detect the minimum titer of virus was 10- 4. 0 / mL. The primers and PVM plasmids stored at -30 ℃ refrigerator for 12 months were used for PCR detection, and the target fragment of about 249 bp could still be amplified. The lung tissue samples test result of 27 mice, 9 rats and 5 gerbils submitted for routine examination were all negative. The result showed that 7 lung tissue samples of the 19 mice infected with PVM were positive. Among the 8 ICLAS international comparison samples tested, 1 sample was positive for PVM nucleic acid, which was in line with the expected result.Conclusion The established PVM RT-PCR method is good in specificity, sensitivity, repeatability and stability,and can be used for the monitoring of mice, rats, gerbils and other experimental animals and the detection of related samples.

Key words:

pneumonia virus of mice, RT-PCR, experimental animals, application