Study on enrichment method of SARS-CoV-2 in sewage based on magnetic beads and ultrafiltration
Objective To develop a rapid and effective method to enrich and concentrate novel coronavirus in sewage,and provide the technical support for the monitoring of SARS-CoV-2 in urban sewage and epidemic prevention and control.Methods The liquid sample microbial collection and enrichment instrument(LM Capturer-5)was used to enrich the novel coronavirus in the spiked liquid samples.RT-qPCR was used to detect the concentration of SARS-CoV-2 nucleic acid before and after enrichment.The recovery rate was calculated to assess the enrichment efficiency of the method for the virus in liquid samples.Additionally,the enrichment efficiency of the LM Capturer-5 method was compared with that of the polyethylene glycol precipitation method.Results After enrichment,the Ct values of the clean samples decreased by 3-5,while the Ct values of the sewage samples decreased by 1-4.The recovery rate of the method established in this study for enriching SARS-CoV-2 in sewage ranged from 13.50％to 44.20％,while the recovery rate of the polyethylene glycol precipitation method ranged from 8.83％to 38.85％.There was no significant difference between the two methods(P＞0.05).The liquid sample microbial collection and enrichment instrument(LM Capturer-5)was used to enrich SARS-CoV-2 in 60 wastewater samples.After enrichment,the positive rate detected was 63.33％,which was consistent with the standard method.The spolyethylene glycol precipitation method was used for consistency test as the standard method,and the Kappa coefficient was 0.785,indicating medium to high consistency.Conclusion This method has a good enrichment effect for SARS-CoV-2 in sewage,and is simple to operate and can be automated.It provides an additional technical reserves for the monitoring of SARS-CoV-2 in the sewage in China,and provides help for the enrichment and detection of multiple pathogenic microorganisms in liquid samples.
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