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Development and characterization of a digital CRISPR/Cas13a based assay for rapid and sensitive diagnosis of severe fever with thrombocytopenia syndrome virus

Sensors and Actuators: B. Chemical. 2023-05; 
Lulu Zuo , Jing Miao , Dongmei He , Zhixin Fang , Xin Zhang , Changyun Sun , Xiaoling Deng , Xiaolin Ma , Chao Yu , Gary Wong
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DNA Sequencing Briefly, the segment S of SFTSV F genotype (AH12 strain, GenBank accession HQ141591.1) with appended upstream T7 promoter sequences (5′-AAATTAATACGACTCACTATAGGG-3′) was constructed commercially (GenScript Biotech Corporation), and then transcribed in vitro using the HiScribe T7 High Yield RNA Synthesis Kit (NEB), following manufacturer instructions. /For specificity tests, the full-length S segment of four Crimean-Congo hemorrhagic fever virus (CCHFV) genotypes (Africa II: Congo 3010 strain; Europe I: Drosdov strain; Asia II: Baghdad 12 strain and Asia I: CLT/T105146 strain) and full-length N gene of Nipah virus (NiV) were constructed commercially (GenScript Biotech Corporation). /The crRNA sequence with appended upstream T7 promoter sequences was constructed commercially (GenScript Biotech Corporation) and then transcribed in vitro using the HiScribe T7 High Yield RNA Synthesis Kit (NEB), following manufacturer instructions. Get A Quote
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摘要

Severe fever with thrombocytopenia syndrome virus (SFTSV) is a re-emerging pathogen causing sporadic outbreaks in China, Japan, and Korea with ortality rates of up to 30 %. There are currently no licensed vaccines or therapeutics, and early diagnosis of infected cases is critical for outbreak control and prevention. Assays based on isothermal nucleic acid amplification and CRISPR/Cas have recently been developed against several pathogens without the need for specialized laboratory equipment. However, these assays are qualitative in nature and cannot provide quantitative results. Here, we developed three candidate diagnostic assays for field or clinical detection of SFTSV infections. The two-step CRISPR/Cas13a... More

关键词

Severe fever with thrombocytopenia syndrome virus Two-step CRISPR/Cas13a One-pot CRISPR/Cas13a Digital CRISPR/Cas13a based assay Quantitative Point-of-care testing
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