豬瘟病毒封套醣蛋白Erns之特異性單株抗體CW813及其於間接三明治ELISA抗體檢測之應用SPECIFIC MONOCLONAL ANTIBODY CW813 AGAINST CLASSICAL SWINE FEVER VIRUS GLYCOPROTEIN ERNS AND ITS APPLICATION TO AN INDIRECT SANDWICH ELISA

專利類型

發明

專利國別 (專利申請國家)

中華民國

專利申請案號

100147899

專利證號

I 490229

專利獲證名稱

豬瘟病毒封套醣蛋白Erns之特異性單株抗體CW813及其於間接三明治ELISA抗體檢測之應用SPECIFIC MONOCLONAL ANTIBODY CW813 AGAINST CLASSICAL SWINE FEVER VIRUS GLYCOPROTEIN ERNS AND ITS APPLICATION TO AN INDIRECT SANDWICH ELISA

專利所屬機關 (申請機關)

國立中興大學

獲證日期

2015/07/01

技術說明

豬瘟病毒(classical swine fever virus, CSFV) 封套醣蛋白Erns是具有RNA降解酵素活性的病毒蛋白。以Erns建構之間接三明治型酵素連結免疫吸附法(indirect sandwich ELISA)可做為區別診斷豬隻體內抗體來源之檢驗方法。本研究之目的為製備生產能夠專一性辨識Erns之單株抗體CW813，此抗體能辨識病毒表現之Erns蛋白及酵母菌表現之Erns (yErns)。利用單株抗體CW813及yErns製備成抗原盤所建立之間接三明治ELISA抗體檢測方法顯示良好之特異性與敏感性， 25個陰性血清皆呈現陰性反應，特異性為100%；而29個陽性血清則檢出率可達96.6% (28/29)。此方式建立之ELISA檢測方式可有效區別不同來源的豬隻血清且具有操作簡單及降低成本之優點。 Classical swine fever virus (CSFV) Erns is an envelope glycoprotein possessing RNase activity. The Erns-based enzyme-linked immunosorbent assay (ELISA) has been considered a discriminating diagnostic test for differentiating infected from vaccinated animals (DIVA). The purpose of this study was to produce a specific monoclonal antibody (MAb) to Erns for further developing an indirect sandwich ELISA. The MAb CW813 was shown to specifically recognize the Pichia pastoris yeast-expressed Erns (yErns). The reactivities of swine serum against yErns using diagnostic ELISA showed 100% of specificity. The detected serum was positive in serum neutralizing test were 96.6% of sensitivity. Furthermore, MAb CW813 was utilized as a capture antibody to develop a yErns-based indirect sandwich ELISA for detecting swine antibody to Erns. The assay demonstrated a high sensitivity and specificity that may provide an alternative method for developing a diagnostic kit with easy manipulation and low cost.

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