CRISPR/Cas12a 技术结合免疫层析条用于非洲猪瘟病毒的便携式检测。

CRISPR/Cas12a technology combined with immunochromatographic strips for portable detection of African swine fever virus.

机构信息

Institute for Brain Research and Rehabilitation, Guangdong Key Laboratory of Mental Health and Cognitive Science, Center for Studies of Psychological Application, South China Normal University, Guangzhou, 510631, China.

Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, China.

出版信息

Commun Biol. 2020 Feb 11;3(1):62. doi: 10.1038/s42003-020-0796-5.

DOI:10.1038/s42003-020-0796-5

PMID:32047240

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7012833/

Abstract

African swine fever virus (ASFV), the aetiological agent of African swine fever (ASF), causes lethal haemorrhagic fever in domestic pigs with high mortality and morbidity and has devastating consequences on the global swine industry. On-site rapid and sensitive detection of ASFV is key to the timely implementation of control. In this study, we developed a rapid, sensitive and instrument-free ASFV detection method based on CRISPR/Cas12a technology and lateral flow detection (named CRISPR/Cas12a-LFD). The limit of detection of CRISPR/Cas12a-LFD is 20 copies of ASFV genomic DNA per reaction, and the detection process can be completed in an hour. The assay showed no cross-reactivity with other swine DNA viruses, and has 100% agreement with real-time PCR detection of ASFV in 149 clinical samples. Overall, the CRISPR/Cas12a-LFD method provides a novel alternative for the portable, simple, sensitive, and specific detection of ASFV and may contribute to the prevention and control of ASF outbreaks.

摘要

非洲猪瘟病毒（ASFV）是非洲猪瘟（ASF）的病原体，可引起家猪致命性出血性发热，死亡率和发病率均很高，并对全球养猪业造成严重后果。现场快速灵敏检测 ASFV 是及时实施控制的关键。在本研究中，我们基于 CRISPR/Cas12a 技术和侧流检测开发了一种快速、灵敏且无仪器的 ASFV 检测方法（命名为 CRISPR/Cas12a-LFD）。CRISPR/Cas12a-LFD 的检测限为每个反应 20 个 ASFV 基因组 DNA 拷贝，检测过程可在 1 小时内完成。该检测方法与其他猪 DNA 病毒无交叉反应性，与 149 份临床样本的实时 PCR 检测具有 100%的一致性。总体而言，CRISPR/Cas12a-LFD 方法为 ASFV 的便携、简单、灵敏和特异性检测提供了一种新的选择，可能有助于预防和控制 ASF 的爆发。