Detection of eight imported viruses using recombinase aided isothermal amplification combined with CRISPR-Cas13a
10.3760/cma.j.cn112866-20211229-00215
- VernacularTitle:重组酶介导的等温扩增技术结合CRISPR-Cas13a蛋白检测8种境外输入性病毒
- Author:
Yue GUO
1
;
Bailin AN
;
Dandan LIU
;
Junhong LUO
;
Kangchen ZHAO
;
Xiaojuan ZHU
;
Yiyue GE
;
Hongbin WU
;
Lunbiao CUI
Author Information
1. 南京医科大学，南京 210029
- Keywords:
Imported infectious disease;
Virus;
Recombinase aided amplification (RAA);
CRISPR-Cas13a
- From:
Chinese Journal of Experimental and Clinical Virology
2022;36(3):245-251
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a rapid, sensitive and specific detection method for important imported viruses based on the recombinase aided amplification (RAA) method and clustered regularly interspaced short palindromic repeats-associated protein 13a (CRISPR-Cas13a) system.Methods:In this study, we selected Japanese encephalitis virus (JEV), Yellow fever virus (YEV), West Nile virus (WNV), Middle East respiratory syndrome coronavirus (MERS-CoV)、Ebola virus (EBOV), Dengue virus (DENV), Rift Valley fever virus (RVFV), Zika virus (ZIKV) as subjects, and designed specific RAA primers and CRISPR RNA(crRNA). The sensitivity and specificity of the method were evaluated. We detected suspected clinical samples of dengue fever and compared with the fluorescent reverse transcriptase-polymerase chain reaction (RT-PCR) technology. Clinical simulation samples of the remaining seven viruses were also detected.Results:The RAA method combined with CRISPR-Cas13a can detect eight pathogens within 40-52 min at 39 ℃. The sensitivity was 1-10 copies/μl. There was no cross-reaction among eight viruses and all clinical samples could be detected by this method.Conclusions:The established RAA combined with CRISPR-Cas13a detection method can sensitively, specifically and quickly detect eight imported infectious disease pathogens.
