| Objective:The EBV-DNA load and EB virus capsid antigen(VCA),nuclear antigen-1(EBNA-1),nuclear antigen-2(EBNA-2),latent membrane protein-1(LMP-1),glycoprotein 350(gp350)and other viral protein expression in serum exosomes of patients with Epstein-Barr virus-associated infectious mononucleosislevels(EBV-IM)were investigated,which will provide some new ideas for the pathogenesis of EBV-IM.Methods:Eighteen patients with EBV-CA-IgM(+)and clinically diagnosed EBV-IM were selected in our hospital as experimental group;Thirteen patients with EBV-CA-IgM(-)/EBV-CA-IgG(+)and clinically diagnosed non-IM were included in the control group.The sera of peripheral blood of the two groups were collected.The exosomes in the sera of the above two groups were separated by differential centrifugation,and the exosomes were identified by Western blot and transmission electron microscopy(TEM).EBV-DNA load in serum exosomes was detected by fluorescence quantitative PCR(FQ-PCR).The expression levels of EBV-CA,EBNA-1,EBNA-2,LMP-1,gp350and other viral proteins in serum exosomes were detected by enzyme-linked immunosorbent assay(ELISA).The experimental data were analyzed using SPSS 20.0 statistical software and related statistical methods.Result:(1)In this experiment,the exosomes in serum of two groups of subjects were successfully isolated.The exosomes were observed by transmission electron microscopy,and the vesicles were in the shape of cups or discs with a diameter of about 30~150 nm.The expression of exosome marker protein CD63 was detected by Western blot.(2)The results of EBV-DNA load in serum exosomes showed that the number of subjects in the experimental group was 18,the EBV-DNA load detection rate was 55.56%,and the number of subjects in the control group was 13,which was not detected.The detection rate of EBV-DNA load in the experimental group was significantly higher than control group,and the difference was statistically significant(?~2=10.661,P<0.05).(3)The expression levels of EBV-CA,EBNA-1,EBNA-2,LMP-1,gp350 and other viral proteins in serum exosomes of the two groups were as follows:the expression levels of EBV-CA,EBNA-1,EBNA-2,LMP-1,gp350 and other viral proteins in the serum exosomes of the experimental group were shown by median(lower quartile,upper quartile)of 0.21055(0,0.45925),0.4541(0,0.49605),0.2157(0,0.465525),0.4442(0,0.484425),0(0,0.451425).The control group was0.59(0.0364,0.6099),0.0052(0,0.55035),0.5983(0.04935,0.63195),0.5236(0.04195,0.56975),0.5837(0.0292,0.6108).There were statistically significant differences in the expression levels of EBV-CA,EBNA-2,LMP-1,gp350 and other viral proteins between the two groups(P<0.05).There was no significant difference in the expression level of EBNA-1 between the two groups(P>0.05).Conclusion:(1)Detection of EBV-DNA load in serum exosomes may have some value in the diagnosis of EBV-IM.(2)The EB virus related components in the serum exosomes of the experimental group and the control group were significantly different,and their significance needs further study. |
PDF Full Text Request