Abstract: To develop a duplex PCR detection kit for gosling plague(GP) and gosling new type viral enteritis (NGVE), and observe the effects of clinical appliance. The gene sequences of gosling plague virus(GPV) and gosling new type viral enteritis virus (NGVEV) was chosen to design the species-specific primers. The PCR amplification conditions were optimized and series of tests were conducted respectively in the specificity, sensitivity, stability, reproducibility, and stored period of the kit, and expanded clinical trials were also carried out. This kit could specifically amplify the expected 375 bp DNA fragment of GPV virus and 223 bp DNA fragment of NGVEV. There was no cross-reaction with other virus, such as DPV, GPMV, NGVEV and GPV. It was able to detect the presence of GPV as low as 10⁴ELD₅₀/mL and NGVEV as low as 2.5×10^-1ELD₅₀. The test results of different batches of kits of the same sample were consistent. The kit worked well after being stored at room temperature for at least six months. Its effect of clinical application was significant. The results turned out that this duplex PCR detection kit could provide a means for early diagnosis, epidemiological investigation and quarantine of GP and NGVE.

Key words: GP; NGVE; PCR detection kit; development; application