Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China.
Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics, Ministry of Agriculture, Shanghai 200241, China.
Acta Biochim Biophys Sin (Shanghai). 2020 Dec 29;52(12):1413-1419. doi: 10.1093/abbs/gmaa135.
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.
中国首例非洲猪瘟（ASF）疫情于 2018 年在沈阳郊区的一个养猪场报告。此后，ASF 的迅速传播和蔓延已成为养猪业面临的最严重威胁。因此，快速准确地检测非洲猪瘟病毒（ASFV）对于制定有效的疾病控制策略至关重要。在本研究中，我们开发了一种基于 DNA 内切酶靶向 CRISPR 转报告（DETECTR）检测的快速准确的 ASFV 检测方法。通过结合重组酶聚合酶扩增与 CRISPR-Cas12a 蛋白，DETECTR 检测法的最小检测限为 8 个拷贝，与其他猪病毒无交叉反应。临床血液样本通过 DETECTR 检测法检测，与实时聚合酶链反应检测法的符合率为 100%（30/30）。ASFV 的快速准确检测可以促进及时采取根除措施和严格的卫生程序，以控制和预防 ASF 的传播。
