Rapid and accurate detection of African swine fever virus by DNA endonuclease-targeted CRISPR trans reporter assay
Zongjie Li, Jianchao Wei, Di Di, Xin Wang, Chenxi Li, Beibei Li, Yafeng Qiu, Ke Liu, Feng Gu, Minglong Tong, Shuiming Wang, Xiaodong Wu, Zhiyong Ma
Abstract
The first case of African swine fever (ASF) outbreak in China was reported in a suburban pig farm in Shenyang in 2018. Since then, the rapid spread and extension of ASF has become the most serious threat for the swine industry. Therefore, rapid and accurate detection of African swine fever virus (ASFV) is essential to provide effective strategies to control the disease. In this study, we developed a rapid and accurate ASFV-detection method based on the DNA endonuclease-targeted CRISPR trans reporter (DETECTR) assay. By combining recombinase polymerase amplification with CRISPR-Cas12a proteins, the DETECTR assay demonstrated a minimum detection limit of eight copies with no cross reactivity with other swine viruses. Clinical blood samples were detected by DETECTR assay and showed 100% (30/30) agreement with real-time polymerase chain reaction assay. The rapid and accurate detection of ASFV may facilitate timely eradication measures and strict sanitary procedures to control and prevent the spread of ASF.