Abstract
The African swine fever virus (ASFV) is a dsDNA virus that can cause serious, highly infectious, and fatal diseases in wild boars and domestic pigs. The ASFV has brought enormous economic loss to many countries, and no effective vaccine or treatment for the ASFV is currently available. Therefore, the on-site rapid and accurate detection of the ASFV is key to the timely implementation of control. The RNA-guided, RNA-targeting CRISPR effector CRISPR-associated 13 (Cas13a; previously known as C2c2) exhibits a “collateral effect” of promiscuous RNase activity upon the target recognition. The collateral cleavage activity of LwCas13a is activated to degrade the non-targeted RNA, when the crRNA of LwCas13a binds to the target RNA. In this study, we developed a rapid and sensitive ASFV detection method based on the collateral cleavage activity of LwCas13a, which combines recombinase-aided amplification (RAA) and a lateral flow strip (named CRISPR/Cas13a-LFD). The method was an isothermal detection at 37 °C, and the detection can be used for visual readout. The detection limit of the CRISPR/Cas13a-LFD was 101 copies/µL of p72 gene per reaction, and the detection process can be completed within an hour. The assay showed no cross-reactivity to eight other swine viruses, including classical swine fever virus (CSFV), and has a 100% coincidence rate with real-time PCR detection of the ASFV in 83 clinical samples. Overall, this method is sensitive, specific, and practicable onsite for the ASFV detection, showing a great application potential for monitoring the ASFV in the field.
Highlights
Introduction published maps and institutional affilAfrican swine fever (ASF) is a severe infectious disease of swine caused by the African swine fever virus (ASFV), which can have a devastating impact on the pig industry [1,2].The ASFV is a large double-stranded DNA virus belonging to the family Asfarviridae [3].The viral genome comprises between 170 and 190 kilobases that encode more than 150 ORFs (Open Reading Frame) and approximately 165 viral proteins [4,5]
The results showed that all the lateral flow strips of eight swine pathogens results showed that all the lateral flow strips of eight swine pathogens were negative bands, negative bands, while the positive band was exclusively observed on the lateral while the positive band was exclusively observed on the lateral flow strip of the ASFV
Due to the lack of effective vaccine and treatment, ASF control strategies largely depend on the rapid detection and slaughter of infected pigs
Summary
Introduction published maps and institutional affilAfrican swine fever (ASF) is a severe infectious disease of swine caused by the African swine fever virus (ASFV), which can have a devastating impact on the pig industry [1,2].The ASFV is a large double-stranded DNA virus belonging to the family Asfarviridae [3].The viral genome comprises between 170 and 190 kilobases that encode more than 150 ORFs (Open Reading Frame) and approximately 165 viral proteins [4,5]. African swine fever (ASF) is a severe infectious disease of swine caused by the African swine fever virus (ASFV), which can have a devastating impact on the pig industry [1,2]. The viral capsid protein p72, encoded by the B646L gene, is highly conserved and well characterized, making it a widely used target for both nucleic acid detection and phylogenetic analysis [6,7]. ASF has been spreading rapidly in China ever since its emergence in 2018, severely destroying the Chinese swine industry and food safety [8]. ASF affects domesticated and wild pigs of all breeds and ages, with a high mortality rate of nearly 100%. ASF presents with high fever, cyanosis of the skin, and severe bleeding in the lymph nodes [9]. There is no effective vaccine or treatment against ASF yet, so the prevention and control of African iations
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
