Development of influenza A and B viruses detection method based on LAMP-CRISPR-Cas12b-lateral chromatography
Objective To develop a diagnostic method for the rapid detection of influenza A and B viruses in epidemiological investigation site and primary healthcare institutions.Methods A method based on LAMP-CRISPR-Cas12b-lateral chromatography was established,which combined isothermal amplification technology with CRISPR-Cas12b system with Lateral Flow Immunoassay technology to detect influenza A and B viruses.Specific LAMP primer sets and crRNAs for influenza A and B viruses were designed.The sensitivity of the method was verified by using the gradient dilution method while the specificity was verified by using four other respiratory viruses.The method was applied for analysis of 52 ILI surveillance samples to verify practicality.Results A method for the detection of influenza A and B viruses by LAMP-CRISPR-Casl2b-side Lateral Flow Immunoassay was successfully established,with a lower detection limit of 10 copies/μL.The method was highly specific and had no cross-reactivity with the other 4 respiratory viruses.The results of 52 ILI surveillance samples were in good agreement and consistency(kappa=1)with qRT-PCR.Conclusions A method suitable for rapid detection of influenza viruses at the epidemiological investigation site and in primary healthcare institute was established.
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