Observation On The Neutralization Of Antibody To Hepatitis C Virus Antigen And Nucleic Acid And Exploration Of Possible Mechanisms

Objective To investigate whether hepatitis C antibody can inhibit hepatitis C virus antigen and nucleic acid,and provide a new idea for the treatment and prevention of hepatitis C.Methods A total of 48 serum samples from patients with hepatitis C(both RNA and antigen positive,and antibody negative),23 serum samples with hepatitis C antibody positive(both RNA and antigen negative)and 50 serum samples without hepatitis C markers(all of RNA.antigen positive and antibody negative)was collected randomly to complete this experiment,where in the 50 negative serum samples were mixed into one negative serum sample for use in this experiment.The experiment consisted of three parts:HCV antibody absorption test.HCV antigen inhibition test and HCV RNA inhibition test.HCV antibody absorption test:in order to observe the results of the experiment,antibody positive serum was diluted 10 times(critical value of positive antibody test)at the beginning of the experiment.Test group:100μL 10 times diluted antibody positive serum+100μL antigenic positive serum;and Control group:100μL 10 times diluted antibody positive serum+100μL saline.The samples were mixed and put in a water bath of 37℃ for 30 mins,then the automatic enzyme exemption analyzer was used for detection.The antibody content in the sample was determined by ELISA(double antigen sandwich method).The results were expressed by OD values and were proportional to antibody content.HCV antigen inhibition test:Test group:200μL antigen positive serum+200μL antibody positive serum;and Control group:200μL antigen positive serum+200μL saline.The samples were mixed and put in a water bath of 37 C for 30 mins,then the immunobiochemical analyzer was used for detection.The content of antigen in the sample was determined by electrochemiluminescence.HCV RNA inhibition test:set up test group 1.test group 2 and:control group.Test group 1:200μL RNA positive(antigen positive)serum+200μL antibody positive serum+200μL negative serum;Test group 2:200μL RNA positive(antigen positive)serum+200μL antibody positive serum+200μL saline;and Control group:200μL RNA positive(antigen positive)serum+200μL saline+200μL saline.The samples were mixed and put in a water bath of 37 C for 30 mins.The content of RNA in samples was determined by fluorescence quantitative PCR and TMA(transcription mediated amplification).The content of RNA was measured by PCR,and the results were expressed by CT,which was inversely proportional to the content of RNA.In TMA test,the antigenic positive serum was a 10 times ratio diluted sample.The results of the TMA method were expressed by OD values and were proportional to RNA content.Results HCV antibody absorption test:the mean values of R(test group/control group)were calculated,all of which were less than 1 and P<0.05,and significant difference exists.HCV antigen inhibition test:the mean values of R were less than 1 and P<0.05,the difference was statistically significant.HCV RNA inhibition test:results of fluorescent quantitative PCR:the mean values of R(test group 2/control group)were less than 1 and the mean values of 4 groups of R(test group 1/control group)were more than 1.Results of TMA:the results were calculated using a formula S=T107+2 × T108+T109(T is the result of the TMA method,i.e.,OD value),and the calculated results were compared.Compared with control group,there were significant differences between group 1 and control group and between group 2 and control group(P<0.05).There was also significant difference when comparing test group 1 and test group 2(P<0.05).Comparison of mean values of S:Smean value of test group 1>Smean value of test group 2,Smean value of test group 1<Scontrol group,Smean value of test group2<Scontrol group.Conclusion Hepatitis C antibody has certain inhibition effects on antigens and RNA,substances in negative serum capable of inhibiting the effect of hepatitis C antibodies may exist.