Development of A Super-Sensitive Diagnostic Method for African Swine Fever Using CRISPR Techniques

查看全文 作　　者：Meishen [1,2]Ren;Hong [1,2]Mei;Ming [1,2]Zhou;Zhen [1,2]F.Fu;Heyou [1,3]Han;Dingren [1,2]Bi;Fuhu [4]Peng;Ling [1,2]Zhao

机构地区：[1]State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China;[2]Key Laboratory of Preventive Veterinary Medicine in Hubei Province,College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China;[3]College of Science,Huazhong Agricultural University,Wuhan 430070,China;[4]Hubei Center for Animal Disease Control and Prevention,Wuhan 430070,China

出　　处：《Virologica Sinica》索引2021年第36卷第2期,共11页

基　　金：supported by the National Natural Science Foundation of China(31522057 and 31872451 to LZ,31720103917 and 31872452 to ZF)。 摘　　要：African swine fever(ASF)is an infectious disease caused by African swine fever virus(ASFV)with clinical symptoms of high fever,hemorrhages and high mortality rate,posing a threat to the global swine industry and food security.Quarantine and control of ASFV is crucial for preventing swine industry from ASFV infection.In this study,a recombinase polymerase amplification(RPA)-CRISPR-based nucleic acid detection method was developed for diagnosing ASF.As a highly sensitive method,RPA-CRISPR can detect even a single copy of ASFV plasmid and genomic DNA by determining fluorescence signal induced by collateral cleavage of CRISPR-lw Cas13 a(previously known as C2c2)through quantitative real-time PCR(q PCR)and has the same or even higher sensitivity than the traditional q PCR method.A lateral flow strip was developed and used in combination with RPA-CRISPR for ASFV detection with the same level of sensitivity of Taq Man q PCR.Likewise,RPA-CRISPR is capable of distinguishing ASFV genomic DNA from viral DNA/RNA of other porcine viruses without any cross-reactivity.This diagnostic method is also available for diagnosing ASFV clinical DNA samples with coincidence rate of 100%for both ASFV positive and negative samples.RPA-CRISPR has great potential for clinical quarantine of ASFV in swine industry and food security. 关键词：African swine fever(ASFV) Recombinase polymerase amplification(RPA) CRISPR-lwCas13a Sensitive diagnosis

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Development of A Super-Sensitive Diagnostic Method for African Swine Fever Using CRISPR Techniques

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