Establishment of a Detection Method for Areca Palm Velarivirus 1 that can Reduce the False Negative of PCR
To establish a detection method for areca palm velarivirus 1(APV1)that can reduce the false negative of PCR.Referring to the reported conserved genes of Areca catechu primers,the Areca catechu internal standard primers and APV1 primers were designed,three pairs of Areca catechu internal standard primers and two pairs of APV1 primers were synthesized,and PCR amplification was performed to screen Areca catechu internal standard primers and APV1 primers.The duplex PCR amplification method was used to optimize the reaction parameters.The optimal primer concentration ratio of internal standard primer and APV1 primer is 1:4,the final concentration of internal standard primer is 0.2 μmol/L,the final concentration of APV1 primer is 0.8 μmol/L,and the optimal annealing temperature is 54 ℃,the optimum number of cycles is 30 times.The sensitivity results showed that the target band could still be amplified when the concentration of Areca catechu cDNA was diluted to 10-6.The specific results showed that only Areca catechu could detect the target band amplified by the internal standard and APV1.Using the established duplex PCR detection method,40 samples of Areca catechu were tested from Wanning City,Hainan Province,and 25 samples were found to be infected with APV1.The results of this study can provide a the-oretical basis for reducing the false negative of areca palm velarivirus 1 PCR.
NETL
NSTL
万方数据
维普
