The Study Of Making Epstein-Barr Virus IgA-VCA And IgA-EA Reagent Of Cell Immunoenzymatic Assay And Comparing With Enzyme-linked Immunosorbent Assay

Objective Preparation cell immunoenzymatic assay test kit to detect Serum Epstein-Barr Virus IgA-VCA and IgA-EA.evaluate the value of Epstein-Barr virus IgA-VCA and IgA-EA tests using immunoenzymatic assay test kit for serological diagnosis of nasopharyngeal carcinoma(NPC)by compared with enzyme-linked immunosorbent assay(ELISA)test kit.Methods Raise B_95-8cell and Raji cell and trypan blue exclusion experiment were used to observe cell survival,B_95-8cell and Raji cell as target antigen of EB virus VCA and EA and painting on glass carrier as biology slide,and comparing express rate of the biology slide of Shanghai Institute of Biological Products.then preparation peroxidase-labelled anti-human IgA antibody and chromogen/substrate solution,the optimization working concentration of peroxidase-labelled anti-human IgA antibody was determined by square matrix method,which make up kit of immunoenzymatic assay.One hundred and seventy-seven sera of patients with NPC and ninety sera from those who had healthy medical examined were collected.ELISAs for detecting IgA-VCA by Euroimmun kit and Shenzhen kangshengbao kit,and immunoenzymatic assay kit for detecting IgA-VCA were used and compared.For detecting IgA-EA, there are Euroimmun ELISA kit and immunoenzymatic assay kit compared. Results The cultural cell was stained by trypan blue,the survival rate was: B_95-8cell with 96%;Raji cell with 92%.The biology slide of B_95-8cell VCA express rate was 40%,and Raji cell EA express rate was 35%.As compared the biology slide of Shanghai Institute of Biological Products was 41%and 34% respectively.The peroxidase-labelled anti-human IgA antibody optimization working concentration was 1:40.The sensitivity of detecting IgA-VCA using Euroimmun ELISA kit was 67.23%,and the Shenzhen kangshengbao ELISA kit was 64.97%,There was no statistically significant difference between Euroimmun ELISA kit and Shenzhen kangshengbao ELISA kit(McNemar's test for correlated proportions;x²=0.35,P＞0.05),but the sensitivity of detecting IgA-VCA using IEA kit was 92.09%,there was statistically significant difference between IEA kit and Euroimmun ELISA kit(x²=35.85,P＜0.001), the same as between IEA kit and Shenzhen kangshengbao ELISA kit(x²=37.16, P＜0.001).The specificity of detecting IgA-VCA respectively Euroimmun ELISA kit was 95.56%,Shenzhen kangshengbao ELISA kit was 96.67%,IEA kit was 88.89%,there was no statistically significant difference each other(Fisher's exact probabilities,P＞0.05).The sensitivity of detecting IgA-EA using Euroimmun ELISA kit was 58.76%,IEA kit was 68.36%,there was statistically significant difference between Euroimmun IEA kit and ELISA kit(x²=13.47,P＜0.001),while the specificity,Euroimmun ELISA kit was 100%,IEA kit was 98.89%,there was no statistically significant difference(P＞0.05).Conclusion The effect of Making Epstein-Barr virus IgA-VCA and IgA-EA Reagent of Cell Immunoenzymatic Assay is good.The sensitivity of detection of serum Epstein-Barr Virus IgA-VCA and IgA-EA by IEA kit was higher than that of IgA-VCA and IgA-EA using ELISA kit,for the specificity,there was no difference.The sensitivity of detection of serum Epstein-Barr Virus IgA-VCA by IEA was high,but the specificity was low;The sensitivity of detection of serum Epstein-Barr Virus IgA-EA by IEA was low,but the specificity was high. The two IEAs,namely,IgA-VCA and IgA-EA,have complementary effect on serological diagnosis of NPC.