Abstract: In order to detect African swine fever virus (ASFV) quickly by ordinary PCR, In this study, we established a common PCR method for ASFV B646L and F778R dual genes by two pairs of specific primers which were designed based on the target sequences of ASFV B646L and F778R genes respectively. And its specificity, sensitivity and reproducibility were tested. It showed that the optimal combination of PCR system (25.0 μL system) was Pfu enzyme 0.2 μL, 10×Reaction Buffer 2.5 μL, 2.5 mmol/L dNTP Mixture 2.5 μL, B646L upstream primer and B646L downstream primer were 1.5 μL, F778R upstream primer, F778R downstream primer each 0.5 μL, DNA template 1.0 μL, annealing temperature 57℃, annealing time 30 s. The method was only used for specific amplification of ASFV B646L and F778R genes, without cross-reaction. The lower limits of detection of positive plasmid B646L and F778R were 7.2×107 copies/μL and 1.5×106 copies/μL. The test repeatability is good. This study has important value for improving the accuracy, specificity and efficiency of clinical diagnosis of ASFV, and provides reliable technical support for rapid detection of ASFV.

Key words: African swine fever; multiplex PCR; B646L gene; F778R gene; quick detection

（Acta Laser Biology Sinica, 2023, 32(6): 554-560）