One-pot one-step detection platform for severe fever with thrombocytopenia syndrome virus via the CRISPR/Cas12a detection system.

IF 4 3区 医学 Q2 VIROLOGY
Jiwei Shu, Qilong Tan, Zhe Huang, Tongjie Zhang, Ling Ye, Shuqin Fu, Zhilei Mao
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引用次数: 0

Abstract

Severe fever with thrombocytopenia syndrome virus (SFTSV) is a tick-borne virus that primarily causes SFTS. Although a common testing route is available, a timely, conventional and accurate method for SFTSV detection is urgently needed. In the present study, we established a platform that combines the recombinase polymerase amplification (RPA) assay with the clustered regularly interspaced short palindromic repeats-CRISPR associated proteins (CRISPR/Cas) 12a technique in one step in one pot. The procedure can be completed within 45 min at a constant temperature without a sophisticated instrument. This method targets the S gene of SFTSV, with a detection limit (LoD) of 11.7 copies per reaction and high specificity, without cross reactivity with other pathogens. Furthermore, across 46 test samples, this method achieved 89.13% consistency with the PCR method (41/46). Together, the reaction system developed in the present study provides not only a novel method for SFTSV detection but also an alternative method for detecting RNA viruses.

基于CRISPR/Cas12a检测系统的发热伴血小板减少综合征病毒“一锅一步”检测平台
发热伴血小板减少综合征病毒(SFTSV)是一种蜱传病毒,主要引起发热伴血小板减少综合征。虽然有常用的检测途径,但迫切需要一种及时、常规、准确的SFTSV检测方法。在本研究中,我们建立了一个平台,将重组酶聚合酶扩增(RPA)检测与聚集规律间隔短回文重复序列-CRISPR相关蛋白(CRISPR/Cas) 12a技术在一个锅中一步结合。该过程可以在恒温下45分钟内完成,无需复杂的仪器。该方法以SFTSV S基因为靶点,每次反应的检出限(LoD)为11.7拷贝,特异性高,与其他病原体无交叉反应性。此外,在46个测试样本中,该方法与PCR方法的一致性达到89.13%(41/46)。总之,本研究建立的反应体系不仅为SFTSV的检测提供了一种新的方法,也为RNA病毒的检测提供了一种替代方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Virology Journal
Virology Journal 医学-病毒学
CiteScore
7.40
自引率
2.10%
发文量
186
审稿时长
1 months
期刊介绍: Virology Journal is an open access, peer reviewed journal that considers articles on all aspects of virology, including research on the viruses of animals, plants and microbes. The journal welcomes basic research as well as pre-clinical and clinical studies of novel diagnostic tools, vaccines and anti-viral therapies. The Editorial policy of Virology Journal is to publish all research which is assessed by peer reviewers to be a coherent and sound addition to the scientific literature, and puts less emphasis on interest levels or perceived impact.
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