Shu Jiwei, Tan Qilong, Huang Zhe, Zhang Tongjie, Ye Ling, Fu Shuqin, Mao Zhilei
Department of Medical Laboratory, Disease Prevention and Control Center of Daishan County, Zhoushan, Zhejiang, China.
Department of Infectious Disease Prevention and Control, Disease Prevention and Control Center of Daishan County, Zhoushan, Zhejiang, China.
Virol J. 2025 Jun 24;22(1):203. doi: 10.1186/s12985-025-02772-0.
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.
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（SFTS）。尽管已有常用的检测途径，但仍迫切需要一种及时、传统且准确的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）分析与成簇规律间隔短回文重复序列-CRISPR相关蛋白（CRISPR/Cas）12a技术在一步一锅法中结合起来。该程序可在恒温下45分钟内完成，无需复杂仪器。该方法靶向SFTSV的S基因，检测限为每个反应11.7个拷贝，特异性高，与其他病原体无交叉反应。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）。本研究开发的反应系统不仅为SFTSV检测提供了一种新方法，也为RNA病毒检测提供了一种替代方法。
