LAMP assay coupled with CRISPR/Cas12a system for portable detection of African swine fever virus
文献类型： 外文期刊
作者： Yang, Bo 1 ; Shi, Zhengwang 1 ; Ma, Yuan 1 ; Wang, Lijuan 1 ; Cao, Liyan 1 ; Luo, Juncong 1 ; Wan, Ying 1 ; Song, Rui 1 ; Ya 1 ;
作者机构： 1.Chinese Acad Agr Sci, Lanzhou Vet Res Inst, State Key Lab Vet Etiol Biol, Key Lab Anim Virol,Minist Agr,Natl Foot & Mouth D, Xujiashan 1, Lanzhou 730046, Gansu, Peoples R China
2.Shenzhen Bioeasy Biotechnol Co Ltd, Res & Dev Dept, Shenzhen, Peoples R China
关键词： African swine fever virus (ASFV); CRISPR system; LAMP assay; portable
期刊名称：TRANSBOUNDARY AND EMERGING DISEASES （ 影响因子：5.005； 五年影响因子：4.622 ）
ISSN： 1865-1674
年卷期：
页码：
收录情况： SCI
摘要： African swine fever (ASF) is one of the most severe infectious diseases of pigs. In this study, a loop-mediated isothermal amplification (LAMP) assay coupled with the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system was established in one tube for the detection of the African swine fever virus (ASFV) p72 gene. The single-stranded DNA-fluorophore quencher reporter and CRISPR-derived RNA were screened and selected for the CRISPR detection system. In combination with LAMP amplification assay, the detection limit for the LAMP-CRISPR assay can reach 7 copies/mu l of p72 gene per reaction. Furthermore, this method displays no cross-reactivity with other porcine DNA or RNA viruses. The performance of the LAMP-CRISPR assay was compared with real-time qPCR tests for clinical samples; a good consistency between the LAMP-CRISPR assay and real-time qPCR was observed. The method shed a light on the convenient, portable, low cost, highly sensitive and specific detection of ASFV, demonstrating a great application potential for monitoring on-site ASFV in the field.
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