Establishment of CRISPR/Cas12a-based molecular detection method for monkeypox virus

VernacularTitle:基于RAA-CRISPR/Cas12a的猴痘病毒核酸检测方法的建立
Author: Meihui LUO ¹ ; Li ZHAO ; Changcheng WU ; Roujian LU ; Ruhan A ; Baoying HUANG ; Yao DENG ; Jiao REN ; Huijuan WANG ; Fei YE ; Wen WANG ; Houwen TIAN ; Wenling WANG ; Wenjie TAN
Author Information

1. 中国疾病预防控制中心病毒病预防控制所，北京　102206
Keywords: Monkeypox virus; CRISPR/Cas12a; Nucleic acid detection; Recombinase-aided amplification
From: Chinese Journal of Experimental and Clinical Virology 2023;37(2):193-200
CountryChina
Language:Chinese
Abstract: Objective:To develop a recombinase-aided amplification (RAA)-clustered regularly interspaced short palindromic repeats(CRISPR)/Cas12a-based nucleic acid assay for monkeypox virus with high specificity and sensitivity.Methods:RAA primers and CRISPR RNA (crRNA) were designed based on the known conserved regions of the monkeypox virus gene and synthesized, and specific crRNAs were screened using fluorescence detection. The sensitivity and specificity of the detection system were evaluated.Results:An RAA-CRISPR/Cas12a-based nucleic acid assay for monkeypox virus was developed with a sensitivity of 2.5 copies/reaction and high specificity without cross-reactivity with ectromelia virus and vaccinia virus.Conclusions:An RAA-CRISPR/Cas12a-based nucleic acid assay for monkeypox virus was established, which would provide a powerful tool for efficient, rapid and specific detection of monkeypox virus.