LAMP assay coupled with CRISPR/Cas12a system for portable detection of African swine fever virus

文献类型： 外文期刊

第一作者： Yang, Bo

作者： Yang, Bo;Shi, Zhengwang;Ma, Yuan;Wang, Lijuan;Cao, Liyan;Luo, Juncong;Wan, Ying;Song, Rui;Tian, Hong;Zheng, Haixue;Yang, Bo;Yan, Yiyong;Yuan, Kehu

作者机构：

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

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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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