| Since the novel coronavirus outbreak in 2019,the epidemic has caused great impact on humans worldwide.With the unremitting efforts of researchers,the clinical detection methods of the novel coronavirus are various and have their own strengths.Because the novel coronavirus spreads very quickly and easily causes clustered spread,but the pathogenicity is reduced,the state has formulated a comprehensive liberalization policy for the epidemic situation and encouraged home self-inspection and self-testing.Colloidal gold immunochromatography antigen self-testing methods and reagents meet national policies and social needs.In this paper,a colloidal gold immunochromatographic assay was constructed by labeling the antibody with colloidal gold as a tracer marker based on the principle of double antibody sandwich detection using commercially available novel coronavirus highly conserved nucleocapsid N protein antibodies as raw materials,and the raw materials and detection system were optimized to prepare the novel coronavirus antigen self-testing closed,visualized,and lateral immunochromatographic reagent strips.The sensitivity and specificity of this method were studied,and finally the application detection of the novel coronavirus positive samples was realized.Through the optimization of reaction system for colloidal gold-labeled antibodies with different particle sizes,3 pairs of test line N protein antibody pairing from different sources,4 pairs of control line antibody pairing and NC membrane and other raw materials,closed detection strips were prepared.The lowest detection limit of recombinant N protein reached 5 pg/m L.At the same time,cross-reaction studies were performed on inactivated solutions of adenovirus,respiratory syncytial virus,Staphylococcus aureus and other pathogens.The results showed that there was no cross-reaction to other pathogens.Nucleic acid and antigen were detected in 101 samples,including 56 cases of novel coronavirus nucleic acid positive samples and 45 cases of nucleic acid negative samples.The positive coincidence rate was 96.43%,the negative coincidence rate was 100%,and the total coincidence rate reached 98.02%.The test strip has been validated by recombinant N protein and novel coronavirus samples to meet the requirements of rapid,accurate,on-site testing and home self-testing,with good sensitivity and specificity,coincidence rate higher than the main performance indicators of similar products,and has certain advantages in detection limit and biosafety.As an important detection method for the novel coronavirus infection,this method can effectively solve the problem that nucleic acid detection depends on instruments and equipment and has a long detection time,meet the needs of clinical diagnostic application,and has a wide range of applicability,which is of great significance for actively responding to the orderly liberalization policy of national novel coronavirus epidemic and preventing and controlling the rapid spread of the epidemic. |
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