A RAPID CRISPR/CAS13A SYSTEM-BASED METHOD FOR DETECTING SEVERE FEVER WITH THROMBOCYTOPENIA SYNDROME VIRUS
Journal Title: Acta Parasitologica et Medica Entomologica Sinica - Year 2024, Vol 31, Issue 3
Abstract
[Objective] To develop a rapid and sensitive on-site nucleic acid detection method based on the clustered regularly interspaced short palindromic repeats (CRISPR) system to detect severe fever with thrombocytopenia syndrome virus(SFTSV). [Methods] Firstly, a specific conserved region of the SFTSV S gene was identified by sequence alignment analysis, followed by the design and selection of multienzyme isothermal rapid amplification (MIRA) primers and target CRISPR RNA (crRNA); detection results were read using fluorescence signals and immunochromatographic paper. Finally, in order to increase sensitivity, multiple detection targets of crRNA were incorporated into a single reaction system. The effectiveness of this method for field nucleic acid detection in tick samples was validated using simulated tick-infected samples and field ticks. [Results] The minimum detection limit (MDL) of the MIRA-CRISPR/Cas13a fluorescence assay for SFTSV was 1 copy/μL, while that of the stripe-based method was 10 copies/μL. No cross-reactivity was observed between SFTSV and the other four viral controls, indicating the high specificity of the MIRA-CRISPR/Cas13a system. The method demonstrated superior sensitivity for nucleic acid detection in simulated tick infection samples compared to RT-PCR. The analysis of 29 field tick samples revealed that 20 and 9 samples were positive and negative, respectively (100% agreement with RT-PCR results). [Conclusions] The proposed method could facilitate the rapid detection of SFTSV in the field.
Authors and Affiliations
Kang-hui DING, Jun HUANG, Jiao FAN, Shao-fu QIU, Hong-bo LIU, Hong-bo LIU, Hong-bin SONG
Keywords
Related Articles
- EP ID EP753996
- DOI 10-3969/j.issn.1005-0507-2024-03-004
- Views 7
- Downloads 0
SUBCELLULAR LOCALIZATION OF AMINO ACID TRANSPORTER CPAAT1 IN CRYPTOSPORIDIUM PARVUM
[Objectives] To investigate the subcellular localization of amino acid transporter 1 (CpAAT1) in Cryptosporidium parvum. [Methods] Based on bioinformatics analysis of the amino acid sequence of CpAAT1, specific polypepti...
COMPLETE GENOME OF AEDES ALBOPICTUS ANPHEVIRUS FROM PORT AREAS IN JIANGXI PROVINCE
[Objective] Insect-specific viruses (ISVs) in Aedes albopictus are closely associated with the transmission and replication of pathogenic infectious agents, such as dengue, chikungunya and Zika virus. In this study, thei...
SUBCELLULAR LOCALIZATION AND ADHESION CHARACTERISTICS OF FIBRONECTIN TYPE Ⅲ DOMAIN-CONTAINING PROTEIN IN CRYPTOSPORIDIUM PARVUM
[Objective] This study was performed to investigate the subcellular localization and adhesion properties of the Cryptosporidium parvum fibronectin type Ⅲ domain-containing protein (CpFN3). CpFN3 is a 2 430-aa single-pass...
RESEARCH PROGRESS ON HEDGEHOGS CARRYING ZOONOTIC PATHOGENS
Wild hedgehogs may act as carriers and/or hosts for bacterial, viral, and fungal pathogens with zoonotic potential, posing a significant threat to humans. Humans are increasingly invading hedgehog habitats, disrupting th...
ESTABLISHMENT AND APPLICATION OF A QPCR-BASED METHOD FOR WOLBACHIA GENOTYPE AND DENSITY DETECTION IN AEDES ALBOPICTUS
[Objective] Naturally, Aedes albopictus carries both the A and B supergroups of Wolbachia strains, which are symbiotic bacteria that can influence the transmission of mosquito-borne diseases by inhibiting the ability of...