臺灣飯匙倩神經毒及其抗體的酵素免疫分析

Abstract

利用酵素免疫分析（Enzyme-immunoassay）定義各種抗原和抗體或分析抗原與抗體之間的相互作用（cross-neaction）具有下列幾種優點： 1、 僅需簡單的儀器和設備。 2、 容易操作。 3、 試劑成本低，並具有持久性。 4、 具有高度的特異性和靈敏度。 5、 分析迅速，不費時。 6、 不會造成放射線物品的污染和危害。 因此，酵素免疫分析的日趨普及並且逐漸取代放射線免疫分析（Radioimmunoassay）是其來的目的。在本論文中，我們引進幾種不同型號的分析方法，針對其檢測臺灣飯匙倩神經毒（Formosan cobra Neurotoxin）和飯匙倩蛇毒抗血清的靈敏度做比較和分析。 本實驗採用山葵過氧化酵素（horseradish peroxidase）當作分析的指標，而聚苯乙烯試管（polystyrene tube）則用做為固相（solid phase）以便於分離已結合或是未結合的抗原或是抗體。 首先，我們利用“三明治”（sandwich）方法來幾種交互反應試驗的結果。對於蛇毒的分類，命名的系統化、科學化、合理化，都將是不可或缺的理論根據。

Several sensitive and simple methods of enzyme-immunoassay for the quantitative determination of cobra neurotoxin and anti-cobra venom serum were reported. Horseradish peroxidase has been conjugated to horse immunoglobulin G or goat anti-horse IgG. Such conjugates, prepared by Schiff base formation between the aldehyde group from the periodate-oxidized carbohydrate moiety of peroxidase and the amino group from the IgG, was used in the immunoassay. Disposable polystyrene tubes were used as solid phase. In the sandwich method of assay, tubes coated with IgG were incubated with NTX followed by an enzyme-labeled IgG. The enzyme remaining in the tubes after washing provided a measure of the amount of NTX. When ELISA was used , tubes were coated with NTX and then incubated with horse IgG followed by incubation with enzyme conjugated goat anti-horse IgG．Though satisfactory result was not obtained with sandwich method, sensitivities as high as 10 ng/ml for NTX and 105 fold dilution for antiserum were obtained with the enzyme-linked immunosorbent assay. Nonspecific adsorptions or horse IgG and goat anti-horse IgG were negligible.