文献类型： 外文期刊
作者： Wu, Mengting 1 ; Chen, Mengmeng 1 ; Qiu, Rulong 1 ; Ge, Lei 1 ; Fan, Zhiyu 1 ; Hu, Bo 1 ; Wei, Houjun 1 ; Li, Yiming 1 ; Wang, Fang 1 ; Song, Yanhua 1 ;
作者机构： 1.Jiangsu Acad Agr Sci, Minist Agr, Key Lab Vet Biol Engn & Technol, Inst Vet Med, Nanjing, Peoples R China
2.Nanjing Agr Univ, Coll Vet Med, Nanjing, Peoples R China
3.Guotai Taizhou Ctr Technol Innovat Vet Biol, Taizhou, Jiangsu, Peoples R China
关键词： CRISPR/Cas12a; portable; rabbit hemorrhagic disease virus (RHDV); RT-LAMP
期刊名称：TRANSBOUNDARY AND EMERGING DISEASES （ 影响因子：3.0； 五年影响因子：3.4 ）
ISSN： 1865-1674
年卷期： 2024 年 2024 卷
页码：
收录情况： SCI
摘要： Rabbit hemorrhagic disease is a highly contagious and acute fatal disease caused by rabbit hemorrhagic disease virus (RHDV). The first outbreak of RHDV2 in 2020 has posed a serious threat to the rabbit breeding industry in China. An effective and specific detection strategy for RHDV GI.1 (RHDV1) and GI.2 (RHDV2) is urgently needed. In this study, we established a reverse transcription loop-mediated isothermal amplification (RT-LAMP)-CRISPR/Cas12a-based dual readout portable detection platform. The platform showed excellent specificity to identify RHDV1 and RHDV2 strains and no cross-reaction with other prevalent pathogens of rabbit. The detection limit for RHDV1 and RHDV2 by RT-LAMP-CRISPR/Cas12a could reach 10 copies/mu l of the VP60 gene per reaction. Furthermore, 74 clinical samples were detected for both RHDV1 and RHDV2. RT-LAMP-CRISPR/Cas12a-based dual readout portable detection platform showed 25.68% (19/74) RHDV1-positive samples, 43.24% (32/74) RHDV2-positive samples, and 8.11% (6/74) RHDV1/RHDV2 double positive samples, respectively. The coincidence rates of detection RHDV1 and RHDV2 between RT-LAMP-CRISPR/Cas12a and quantitative real-time-polymerase chain reaction (qPCR) were both 97.30%. RT-LAMP-CRISPR/Cas12a showed higher sensitivity and detection rate compared with qPCR. Moreover, the results were visible to the naked eye within 1.5 h combined with lateral flow strips (LFSs) and visual fluorescence. The RT-LAMP-CRISPR/Cas12a portable platform has the advantages of high sensitivity, specificity, fast, low equipment requirements, which can be used in clinical practice in rural areas and resource-limited settings.
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