Fitra Bp, Nurmadinah (2021) "Pengembangan Deteksi African Swine Fever Berbasis Protein Rekombinan Spesifik P72-Asfv Yang Diinduksi Pada Hewan Coba Kelinci (Oryctolagus Cuniculus) ". Sarjana thesis, Universitas Brawijaya.
Abstract
"African swine fever (ASF) adalah penyakit hemoragik mematikan yang menyerang babi liar dan domestik dengan tingkat penyebaran penyakit yang sangat cepat. Protein p72 merupakan virion penyusun kapsid virus ASF yang menjadi imunogen utama yang ditemukan dalam infeksi alami pada babi. Penelitian ini bertujuan untuk membuktikan bahwa protein rekombinan p72 bersifat imunogenik, serta untuk mengetahui spesifisitas antibodi poliklonal p72 dalam mengenali protein rekombinan p72. Protein rekombinan p72 merupakan protein yang diperoleh dari hasil rekayasa genetik dengan memindahkan gen pengkode protein p72 ke sel bakteri E.coli. Sampel protein rekombinan p72-ASFV yang digunakan berasal dari PUSVETMA Surabaya. Selanjutnya dilakukan purifikasi dan karakterisasi protein rekombinan p72 dengan metode elektroforesis SDS-PAGE dengan melihat profil pita protein p72 yang memiliki berat molekul 72 kDa. Untuk memperoleh antibodi protein p72 dilakukan imunisasi menggunakan kelinci putih New Zealand (Oryctolagus cuniculus) berjenis kelamin jantan umur kurang lebih 6 bulan dengan berat badan 4 kg. Kelinci diimunisasi dengan isolate protein p72 yang diemulsikan dalam adjuvant (1:1) secara subkutan dengan dosis 800 µL. Imunisasi pertama dengan penambahan CFA 400 µL dan IFA 400 µL pada booster pertama dan booster kedua. Evaluasi imunogenesitas protein rekombinan p72 dilakukan dengan metode indirect ELISA sedangkan uji spesifisitas antibodi p72 terhadap antigen p72 dilakukan dengan uji Western Blot. Serum kelinci hasil induksi protein p72 dikoleksi setiap minggu selama 7 minggu. Hasil uji titer antibodi dengan metode indirect ELISA tertinggi pada bleeding minggu ke-7 dengan nilai optical density sebesar 0.337. Hasil uji spesifitas dengan uji Western Blot menunjukkan bahwa antibodi p72 dapat mengenali antigen p72 pada berat molekul 72 kDa.
English Abstract
African swine fever (ASF) is a deadly hemorrhagic disease that attacks wild and domestic pigs with the infection rate is very fast. ASF disease caused by a virus of double-stranded DNA of the genus Asfavirus. Protein p72 is an ASF virus virion capsid composer who became major immunogens that are found in the natural infection in pigs. This study aims to find the candidate ASF detection using rabbit polyclonal antibodies induced by the recombinant protein p72-ASFV. Recombinant p72 protein is a protein obtained from genetic engineering by transferring the gene encoding the protein p72 to E.coli bacterial cells. SampleASFV p72 recombinant protein is derived from PUSVETMA Surabaya. Furthermore, the purification and characterization of recombinant protein p72 by SDS-PAGE electrophoresis method to see the profile of p72 protein band with molecular weight of 72 kDa. To obtain the p72 protein antibody, immunization was carried out using a male New Zealand white rabbit (Oryctolagus cuniculus) aged approximately 6 months and weighing 4 kg. Rabbits were immunized with p72 protein isolate which is emulsified in adjuvant (1: 1) subcutaneously at a dose of 800 mL. The first immunization with CFA addition of 400 mL and 400 mL IFA on the first booster and a second booster. Evaluation of recombinant protein p72 imunogenesitas conducted using indirect ELISA test while specificity antibodies against the antigen p72 p72 done by Western Blot test. Rabbit serum produced by p72 protein was collected every week for 7 weeks. Antibody titer test results with the highest ELISA indirect method on bleeding week 7 with optical density values for 0337. The results of the specificity test using the Western Blot test showed that the p72 antibody could recognize the p72 antigen at a molecular weight of 72 kDa.
Other obstract
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| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | 052113 |
| Uncontrolled Keywords: | protein rekombinan p72, imunisasi, antibodi poliklonal, : Recombinant protein of p72, immunization, polyclonal antibody |
| Subjects: | 600 Technology (Applied sciences) > 636 Animal husbandry |
| Divisions: | Fakultas Kedokteran Hewan > Kedokteran Hewan |
| Depositing User: | Unnamed user with username rizky |
| Date Deposited: | 22 Oct 2021 07:24 |
| Last Modified: | 24 Feb 2022 16:40 |
| URI: | http://repository.ub.ac.id/id/eprint/185080 |
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