Immunochromatographic Strip Using Baculovirus Expression Nucleoprotein to Detect Avian Influenza Antibody

應用桿狀病毒表現核蛋白質以免疫色層分析試片檢測家禽流行性感冒抗體

陳光瑜(Kuang-Yu Chen) ； 鄭益謙(Ivan-Chen Cheng) ； 王金和(Ching-Ho Wang)

《台灣農學會報》 17卷1期 (2016/03) Pp. 76-86

https://doi.org/10.6730/JAAT.201603_17(1).0004

摘要

因禽流感抗體遠比病毒存在雞隻體內時間久，故檢測抗體較檢測病毒較易檢測出陽性雞群，為監測中最常使用的方法之一，因此本研究目的在於建立免疫色層分析試片檢測禽流感抗體。選殖H5N2禽流感病毒之核蛋白質基因，於桿狀病毒系統表現禽流感病毒之核蛋白質，作為抗原進行酵素連結免疫吸附法檢測雞隻禽流感抗體，依血球凝集抑制抗體為金標準，以此重組蛋白質建立之間接型ELISA測試，結果此ELISA敏感度為100%，特異度為96.6%。此外此重組核蛋白質應用於免疫色層分析試片的敏感度為93.2%，特異度為87.5%。結果顯示本禽流感檢測方法可用於檢測至少H5及H6亞型之AI抗體，可以應用於現場快速檢測禽流感抗體。

關鍵字

禽流感抗體 ； 桿狀病毒表現系統 ； 雞 ； 免疫色層分析 ； 核蛋白質

並列摘要

As a part of routine surveillance, detecting avian influenza (AI) antibody has a longer time than detecting AI virus (AIV) itself because the former lasts longer than the latter. In order to detect AI antibody in the field, we developed a membrane-based immunochromatograpic strip (ICS). The nucleoprotein (NP) gene of an H5N2 AIV was cloned and expressed in a baculovirus expression system. The expression NP was applied in an enzyme-linked immunosorbent assay (NP-ELISA) and compared with the hemagglutination inhibition (HI) test. The sensitivity and the specificity of the NP-ELISA based on HI reached 100% and 96.6%, respectively. In addition, the NP was applied in ICS to detect anti-AIV antibodies. The sensitivity and the specificity of NPICS based on HI were 96.5% and 89.1%, respectively. The present NP-ICS detects antibodies at least against H5 and H6 AIVs and provides a quick and convenient method for AIV antibody detection in the field.

並列關鍵字

Avian influenza antibody ； Baculovirus expression system ； Chickens ； Immunochromatography ； Nucleoprotein