Establishment and application of duplex PCR detection system between Anguillid herpesvirus and Eel circovirus

  Objective  The aim of the present study was to establish a duplex PCR method for detection of Anguillid herpesvirus and Eel circovirus.

  Method  Routine PCR primers were preferred to optimize the amplification conditions and test the specificity and sensitivity of the method. At the same time, the sick eel of suspected cases collected from different farm were detected by the duplex PCR method, and compared with the routine PCR method.

  Result  The results showed that the specific fragments of 690 bp and 388 bp were amplified by using the duplex PCR from DNA samples of Anguillid herpesvirus and Eel circovirus, respectively. But the nucleic acid amplification results of Porcine circovirus, Duck circovirus, Koi herpesvirus and Cyprinid herpesvirus were negative. When the positive DNA concentrations of both Anguillid herpesvirus and Eel circovirus were diluted to 5.0×10^-4 μg·mL^-1, the corresponding target fragments could still be detected. 50 samples collected from suspected cases of sick eels were diagnosed by PCR. The infection rate of Anguillid herpesvirus was 16%. The infection rate of Eel circovirus was 68%. The infection rate of Anguillid herpesvirus and Eel circovirus was 8%.

  Conclusion  The results indicate that the duplex PCR detection method has certain specificity and sensitivity, and can be used for the detection and differential diagnosis of recessive cases of clinical infection of Anguillid herpesvirus and Eel circovirus．