Distribution Of Syndrome Factors In Novel Coronavirus Pneumonia And Preparation Of Antibodies Against Virus 3CLpr

Objective: To study the TCM syndrome type,syndrome elements and four diagnosis information of COVID-19 based on literature;to explore the rules of the COVID-19 in order to provide support for better TCM treatment of COVID-19.To facilitate potential pandemic,polyclonal antibody against 3CL protease(3CLpro)of SARS-Co V-2 was generated to provide basis for de novo anti-viral drug development.Methods: 1.China National Knowledge Network(CNKI),WANFANG DATA and VIP were searched from January 2020 to January 2022 to extract TCM syndrome type,disease location,disease syndrome element,information of four diagnoses,number of cases and other information.Excel was used to establish a database,and relevant extracted information was statistically analyzed by SPSS software.2.The 3CLpro coding sequence was amplified by means of PCR and cloned into expression vector p Tri Ex-4 to construct 3CL-p Tr-EX-4,which was transformed into E.coli BL21(DE3)strain and IPTG was used for induction of 3CLpro expression.Recombinant 3CLpro was purified via Ni-NTA resin and used to immune rabbit for antibody production.Western blotting and immunofluorescence assay were used to evaluate the quality of the antibody.Results: 1.Through the collation of literature fifty-three TCM syndrome types of COVID-19 were obtained,among which the major syndromes were epidemic-virus-closed lung syndrome,dampness-heat accumulation at lung syndrome,cold-dampness stagnation of lung syndrome,dampness toxin stagnation of lung syndrome,and deficiency of Qi and Yin syndrome.The most frequent syndromes were lung,exterior,and spleen,and the syndrome was dampness.The most frequent syndromes were fever,cough and fatigue.2.SARS-Co V-2 3CLpro recombinant protein expression vector was successfully constructed,and the 3Clpro was expressed in E.coli and purified.The anti-3CLpro antibodies were obtained by immunization of rabbits with purified 3Clpro protein.Conclusion: 1.The main TCM syndrome types of COVID-19 patients were epidemic virus closed lung syndrome,dampness-heat accumulation of lung syndrome.The symptoms were found mainly in the lung,followed by the exterior and spleen.The disease mainly involves dampness,heat and poison,and the main symptoms were fever,cough and fatigue.2.The antibody against SARS-Co V 3CLpro was successfully obtained which can be used for further anti-viral drug development.