临床输血与检验2025，Vol.27Issue(3)：393-401,9.DOI:10.3969/j.issn.1671-2587.2025.03.018
基于RT-RAA-CRISPR/Cas13a的寨卡病毒检测方法的建立
A CRISPR/Cas13a-Coupled RT-RAA System for Rapid Zika Virus Diagnosis
摘要
Abstract
Objective Zika virus(ZIKV)is an emerging arthropod-borne flavivirus transmitted by Aedes mosquitoes.Recent commentaries regarding ZIKV routes of transmission describe a potential transmission by transfusion.The current approaches in use are low in diagnostic efficiency and coverage area of ZIKV infection.Hence,this study intended to drive the ZIKV infection detection to effectively adaptable for any small to medium-sized laboratory or field survey.Methods We established,optimized,and evaluated a colloidal gold test strip for detection of ZIKV RNA based on CRISPR/Cas13a combined with reverse-transcription recombinase-aided amplification assay(RT-RAA)technology.Results The strip detection of ZIKV RNA was established based on RT-RAA-CRISPR-Cas13a technology with a sensitivity of 4 copies/μL and a specificity of 100%.ZIKV RNA gradient concentration standards and clinical samples were effectively identified by this approach.The positive coincidence rate for ZIKV was 100%,while the negative coincidence rate was 100%.The sensitivity,specificity,positive predictive agreement(PPA)and negative predictive agreement(NPA)values of Zika virus liquid plasma detection were 100%,100%,100%,and 100%,respectively.Conclusion We have developed rapid and portable RT-RAA-CRISPR/Cas13a-based strip of ZIKV RNA detection for patients.This study provides a visual and faster alternative to current PCR-based diagnosis for ZIKA infection.关键词
CRISPR/Cas13a/逆转录重组酶介导链置换核酸扩增/寨卡病毒/即时检测Key words
CRISPR/Cas13a/RT-RAA/ZIKV/point of care test分类
医药卫生引用本文复制引用
张雅路,石耀强,刘金胜,李世林,陈利民,杨春晖..基于RT-RAA-CRISPR/Cas13a的寨卡病毒检测方法的建立[J].临床输血与检验,2025,27(3):393-401,9.基金项目
本课题受四川省自然科学基金项目(No.2025ZNSFSC0678)资助 （No.2025ZNSFSC0678）
