免疫酵素法在豬瘟實驗室診斷上之應用

Laboratory Diagnosis of Hog Cholera by Using Immunoenzymatic Staining Method

魯懿萍(Yi-Pin Lu) ； 潘銘正(Ming-Jeng Pan) ； 傅祖慧(Tsu-Huei Fuh)

《中華民國獸醫學會雜誌》 21卷6期 (1995/12) Pp. 354-364

https://doi.org/10.7009/JTCSVS.199512.0354

摘要

為檢討免疫酵素法在豬瘟實驗室診斷上之應用效果，以直接、間接及Labelled-avidin-biotin（LAB）等3種方法，應用渠草根過氧化氫酶（horseradish peroxidase; HRP）、鹼性磷酸酶（alkaline phosphatase; AP）、半乳糖苷酶（β-galactosidase）等3種酵素，diaminobenzidine（DAB）, aminoethyl carbazole （AEC）, bromochloroindolyl phosphate／nitroblue tetrazolium （BCIP／NBT）, naphthol as-bi phosphate／fast blue (NABP／FB), naphthol as-bi phosphate／new fuchsin（NABP／NF）, bromoch-loroindolyl β-D-galactopyranoside（BCIG; X-Gal）等6種呈色劑，分別檢查接種ALD株豬瘟病毒細胞培養片，及接種同病毒斃死豬扁桃腺冷凍切片中之病毒抗原，同時以直接免疫螢光抗體法做對照。此3種染色方法之結果與對照之直接免疫螢光抗體法者相同，均可檢出豬瘟病毒抗原。其中以使用HRP為標幟酵素，AEC為呈色劑之LAB法效果最佳，可作為臺灣一般實驗室之豬瘟診斷常用法。

關鍵字

免疫酵素法 ； 豬瘟 ； 渠草根過氧化氫酶 ； 鹼性磷酸酶 ； 半乳糖苷酶

並列摘要

This study evaluated the direct, indirect, and labelled-avidin-biotin-(LAB) immunoenzymatic techniques for the laboratory diagnosis of Hog Cholera (HC). The enzymes used for antibody labelling include horseradish peroxidase (HRP), alkaline phosphatase (AP), and ß -galactosidase. And the insoluble substrate (a total of 6) for each enzyme include diaminobenzidine (DAB), aminoethyl carbazole (AEC), bromochloroindolyl phosphate/nitroblue tetrazolium (BCIP/NBT), naphthol as-bi phosphate/ fast blue (NABP/FB), naphthol as-bi phosphate/new fuchsin (NABP/NF), bromocholoroindolyl ß-galactopyranoside (BCIG; X-Gal). ALD strain HC virus infected cells and the tonsil cryosection from experimentally infected pigs were stained by the above methods. AII three immunoenzymatic methods and the direct immunofluorescence staining can successfully detect HC virus antigen in cells. However, LAB method that use HRP-Iabelled antibody and AEC substrate presents the best result.

並列關鍵字

Immunoenzymatic techniques ； Hog cholera ； Horseradish peroxidase ； Alkaline phosphatase ； β-Galatosidase