摘要：Antigen-antibody reaction is an important tool for the analysis of localization of target molecules, including antigenic protein within worm tissues. The purpose of the present research was to demonstrate the ability of immunoglobulin yolk (IgY) anti-excretory/secretory recognized the antigen in the tissue of Ascaridia galli by mean of immunohistochemistry method. The excretory/secretory protein was procured from A. galli and concentrated by mean of vivaspin 30,000 MWCO. IgY was produced by egg yolks of immunized chickens with excretory/secretory, and purified using fast protein liquid chromatography (FPLC) method. A. galli adult worms were cut in transversal and longitudinal section of the center and anterior region. Slides were incubated with both primary IgY for overnight at 4 oC and secondary antibody rabbit anti-chicken IgY HRP-conjugate for one hour at room temperature. The slides were stained with 3-amino, 9-ethylcarbazole (AEC) chromogen, counterstained with Lillie Mayer Haematoxylin, and mounted in glyserin aqueous mount. Antigen-antibody reaction was investigated under a microscope. The result showed that antigen was appeared in the tissues such as cuticle, epicuticle, buccal cavity, and eggs inside the uterine of A. galli. This research concluded that IgY stimulated by the excretory/secretory was able to recognized the antigen scattered in the tissues of A. galli so the IgY could be applied for immunodiagnostic.
其他摘要：Reaksi antigen-antibodi adalah salah satu cara yang penting untuk menganalisis lokasi target molekul, termasuk protein antigen di dalam jaringan cacing. Tujuan riset ini adalah untuk menunjukkan kemampuan immunoglobulin yolk (IgY) anti-ekskretori/sekretori mengenal antigen di dalam jaringan cacing Ascaridia galli melalui metode imunohistokimia. Protein eksretori/sekretori diperoleh dari A. galli dan dipekatkan melalui vivaspin 30.000MWCO. IgY dihasilkan oleh kuning telur dari ayam yang divaksinasi dengan ekskretori/sekretori, dan dimurnikan dengan menggunakan metode fast protein liquid chromatography (FPLC). Cacing A. galli dewasa dipotong secara melintang dan memanjang pada bagian tengah dan atas. Slide dieramkan dengan kedua antibodi, yaitu antibodi primer IgY selama satu malam pada temperatur 4 o C dan antibodi sekunder IgY HRP-conjugate selama satu jam pada temperatur kamar. Slide diwarnai dengan kromogen 3-amino, 9-ethylcarbazole (AEC), dilatar-belakangi oleh Lillie Mayer Haematoxylin, dan direkatkan di dalam gliserin. Reaksi antigen-antibodi diamati di bawah mikroskop. Hasil menunjukkan bahwa antigen ditampilkan di dalam jaringan seperti kutikula, epikutikula, rongga mulut, dan telur di dalam uterus cacing A. galli. Riset ini menyimpulkan bahwa IgY yang dirangsang oleh ekskretori/sekretori mampu mengenal antigen yang tersebar di dalam jaringan A. galli sehingga IgY dapat diaplikasikan untuk imunodiagnostik.
