异源抗原在建立ELISA检测传染性法氏囊病毒抗体中的应用

DEVELOPMENT OF AN ELISA FOR DETECTION OF IBDV ANTIBODY

本文报告采用ｖｅｒｏ细胞增殖的ＩＢＤＶ抗原建立了间接酶联免疫吸附试验（ＥＬＩＳＡ），用于定量检测鸡传染性法氏囊病毒（ＩＢＤＶ）抗体。该法快速、敏感性高、特异性强、重复性好。同时，通过３０份血清样品ＥＬＩＳＡ效价（ＥＴ）的对数值（ｌｏｇＥＴ）与血清Ｐ／Ｎ值（待检血清ＯＤ值与阴性血清ＯＤ值之比）的线性回归分析，得直线方程ｙ＝３．０５８９＋０．０７３９ｘ（ｒ＝０．９１７４），从而血清样品的ＥＴ可通过血清单一稀释度的Ｐ／Ｎ值来计算。用不同来源抗原作ＥＬＩＳＡ表明，从ｖｅｒｏ细胞增殖的抗原比从鸡胚成纤维（ＣＥＦ）细胞增殖的抗原可提高检测血清ＯＤ值近２０％。

The B87 strain of infectious bursal disease virus(IBDV) was multipicated in vero cell and indirect enzyme linked immunosorbent assay(ELISA) for dectection and quantification of IBVD antibody in chickens was developed.A linear relationship was existed between the values of positive/negative(P/N) ratio of serum at 1∶400 and the logarithm of ELISA titer(lgET) determined by a standard serum dilution method of 30 field sera.Regression analysis was used to construct a standard curve and extract a equation from the relationship.The equation was y=3.0589+0.0739x(r=0.9174).The ET of a serum sample could be determined using a single serum dilution by the equation. The result of comparative study showed that the OD values of sera using IBDV antigen from vero cell was increased by about 20% more than using IBDV antigen from chicken embryo fibroblast (CEF) in the indirect ELISA.