基于CRISPR-ERASE技术的现场、高灵敏亨德拉病毒核酸检测方法建立OA

Objective To establish a field-deployable,rapid and highly sensitive detection method for Hendra virus nucleic acidusing clustered regularly interspaced short palindromic repeats(CRISPR)combined with the easy-readout and sensitive enhanced(ERASE)technique on lateral flow test strips.Methods Based on the conserved region of the Hendra virus N gene,specific primers for recombinase-aided amplification(RAA)and CRISPR RNA(crRNA)were designed and synthesized.The target sequence was amplified using RAA technology,followed by detection with the CRISPR/Cas13a system.The amplified products were subsequently analyzed using ERASE lateral flow test strips to interpret the results.Results This assay was able to reliably detect Hendra virus nucleic acid at as low a concentrationas 1 copy/μL within one hour withoutthe use of complex laboratory equipment.There was no cross-reactivity with nucleic acids from seven other RNA viruses that caused similar clinical manifestations.In simulated sample tests,the assay achieved 100%concordance with the expected results.Conclusion This study has established a field-deployable and highly sensitive detection method for Hendra virus nucleic acid based on isothermal amplification combined with the CRISPR-ERASE platform,which can contribute to rapid on-site detection of and control over Hendra virus transmission.