Development and application of a droplet digital polymerase chain reaction (ddPCR) for detection and investigation of African swine fever virus
2018
Wu, Xulong | Xiao, Lü | Lin, Hua | Chen, Shijie | Yang, Miao | An, Wei | Wang, Yin | Yang, Zexiao | Yao, Xueping | Tang, Zizhong
The aim of this study was to develop a droplet digital polymerase chain reaction (ddPCR) method to detect African swine fever virus (ASFV). The methods of ASFV real-time PCR and ddPCR were established and optimal reaction conditions were confirmed. Each method was evaluated for linearity, limit of detection, and specificity. The results indicated that ASFV ddPCR had a high degree of linearity (R2 ≥ 0.998) and specificity. The detection limit was 10 copies/reaction, which was approximately a 10-fold greater sensitivity than real-time PCR. This sensitive method could be used as an efficient molecular biology tool to diagnose ASFV, which is very important for preventing the spread of diseases across borders.
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