家禽流行性感冒病毒抗體免疫色層分析測試片之開發

Abstract

本研究目的在於利用免疫色層分析測試片 (immunochromatographic strip；ICS) 快速檢測及簡單操作的特性，發展針對家禽流行性感冒病毒抗體之檢測方法並運用至現場。實驗中ICS主要利用固定於硝基纖維膜上的抗原，以及帶有膠體金標示的追蹤子 (tracer) 所構成。其中分別以全病毒 (A/chicken/Taiwan/2838V/00 (H6N1)) 及M1重組蛋白質 (recombinant M1；rM1) 做為抗原進行ICS之發展，並使用血球凝集抑制試驗 (hemagglutination inhibition test；HI test) 作為檢測血清之金標準 (gold standard) 進行ICS之評估。全病毒之免疫色層分析測試片 (virus-ICS) 需使用經一次冷凍解凍後之血清進行測試。使用田間血清進行測試時，其敏感性及特異性分別為95.2% (159/167) 及94.3% (150/159)，但其檢測靈敏度低於HI test。至於M1重組蛋白質之免疫色層分析測試片 (rM1-ICS) 方面，rM1無法應用於rM1-ICS以進行田間陽性及陰性血清之區分。總結，本研究發展之virus-ICS仍具有應用於現場快速檢測之潛力。

The purpose of this study was to develop immunochromatographic strips (ICS) for the detection of antibody against avian influenza virus and field application by using rapid procedure and easy operation. In our experiment, ICS was constructed by fixing antigens on the nitrocellulose membrane and the colloidal gold labeled tracer. We separately used the whole virus (A/chicken/Taiwan/2838V/00 (H6N1)) and the recombinant M1 protein (rM1) as the antigens to develop ICSs, using the hemagglutination inhibition test (HI test) as the gold standard for the evaluation of those ICSs. The virus-ICS was applied to the serum pretreated by a freeze-thaw cycle. Although the virus-ICS detection limit was lower than HI test, the sensitivity and specificity reached 95.2% (159/167) and 94.3% (150/159), respectively. On the contrary, the rM1-ICS could not be applied to distinguish the field positive and negative serum samples. In conclusion, virus-ICS could be a potential to be used as rapid test in the field.