Objective: Mink Aleutian disease, viral enteritis and canine distemper are the three major diseases affecting mink health. To establish a multiplex PCR detection method that can detect the three viruses at the same time.
Methods: Three pairs of specific primers were designed for the conserved regions of the three viral genes, and the DNA templates of mink Aleutian virus (ADV) and mink enteritis virus (MEV) and the RNA template of canine distemper virus (CDV) were multiplexed. PCR amplification and optimization of amplification conditions.
RESULTS: PCR could simultaneously amplify specific target bands of 601 bp (ADV), 205 bp (MEV) and 451 bp (CDV). The sensitivity test showed that the minimum nucleic acid detection amount was 2.67×104 copies of ADV per microliter , MEV 3.02×104 copies per microliter and CDV 1.72×105 copies per microliter. The results of clinical samples showed that the results of multiplex PCR and single PCR were consistent.
Conclusion: The established multiplex PCR detection method can rapidly detect ADV, MEV and CDV single or mixed infected clinical samples.