【Animal modeling】-Establishment and preliminary application of FO-PCR detection method for mouse pox virus

  Objective To establish a TaqMan probe-based fluorescent quantitative PCR detection method for mouse pox virus (ECTV), and to detect the nude mole rat tissue samples, mouse tissue samples and chinchilla tissue samples preserved in the laboratory.

  Methods Primer probes were designed according to the crnD gene sequence of mouse pox virus in GenBank, and the FQ-PCR detection method for ECTV was established. The preserved samples were screened, and some positive samples were sequenced and identified.

  Results The established ECTV FQ-PCR detection method has high sensitivity and specificity, and can detect samples with ECTV nucleic acid concentration of 10 copies/μ. The method was used to detect 63 nude mole rat spleen tissue samples and 2 mouse spleen tissue samples in the laboratory, and the results were all negative. After PCR detection, the positive samples were sequenced and compared to ECTV nucleic acid sequences.

  Conclusion It has been verified that the established ECTV FO-PCR detection method can screen ECTV in samples sensitively and specifically, and the potential risk of ECTV contamination should not be ignored in routine monitoring of laboratory mice.