Putri, Novi Aprilia Shania (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
Kombinasi fitobiotik jahe merah (Zingiber officinale var. Rubrum), kunyit (Curcuma longa Linn.), dan temulawak (Curcuma xanthorrhiza Roxb.) berkhasiat sebagai antiinflamasi. TLR2 merupakan reseptor yang terlibat dalam proses inflamasi. IL-6 merupakan mediator inflamasi yang dihasilkan melalui jalur pensinyalan TLR2. Penelitian ini bertujuan untuk melihat interaksi enam senyawa yaitu gingerol-6, paradol-6, kurkumin, bisdemetoksikurkumin, demetoksikurkumin, dan tetrahidrokurkumin terhadap reseptor TLR2 Gallus gallus domesticus menggunakan uji in silico berupa penambatan molekuler serta membuktikan bentuk ikatan penghambatan pada reseptor melalui respon biologis berupa penurunan kadar IL-6 pada serum darah menggunakan uji ELISA. Uji in vivo dilakukan menggunakan Rancangan Acak Lengkap (RAL) terhadap 20 ekor ayam broiler strain cobb slow feather dengan berat rata-rata awal 51,5 gram. Ayam dibagi dalam 5 perlakuan dengan penambahan kombinasi serbuk jahe merah, kunyit, dan temulawak (P1: 0,5%; P2: 1%; P3: 1,5%; P4: 2%; K(-): 0%). Hasil eksplorasi enam senyawa aktif secara in silico menunjukkan bahwa keenam senyawa yang dijadikan ligan mampu berinteraksi dengan reseptor TLR2 Gallus gallus domesticus. Kurkumin memiliki nilai energi ikatan paling rendah ketika berikatan dengan reseptor TLR2 Gallus gallus domesticus dibandingkan dengan kelima ligan uji lain dalam uji in silico dengan skor energi ikatan (ΔG) yaitu -8,0 kkal/mol serta memiliki satu ikatan hidrogen dalam residu asam amino Thr291 pada situs aktif reseptor. Energi ikatan paling rendah dari kompleks ikatan kurkumin dan TLR2 Gallus gallus domesticus membentuk kestabilan energi yang paling baik saat berinteraksi. Hasil uji statistika terhadap kadar IL-6 menggunakan ONE WAY ANOVA dengan tingkat kepercayaan 95% (p<0,05) dan uji lanjutan berupa Tukey test menunjukkan bahwa penambahan kombinasi serbuk jahe merah, kunyit, dan temulawak mampu menurunkan kadar IL-6 secara signifikan pada dosis 1,5% dan 2%. Kesimpulan dari penelitian ini yaitu kombinasi serbuk jahe merah, kunyit, dan temulawak mampu berfungsi sebagai antiinflamasi dilihat dari uji in silico dan in vivo serta dosis optimal penambahan kombinasi serbuk jahe merah, kunyit, dan temulawak sebagai antiinflamasi adalah 1,5%.
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: | Recombinant protein of p72, immunization, polyclonal antibody, protein rekombinan p72, imunisasi, antibodi poliklonal |
| Subjects: | 600 Technology (Applied sciences) > 635 Garden crops (Horticulture) 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 06:23 |
| Last Modified: | 10 Oct 2024 00:11 |
| URI: | http://repository.ub.ac.id/id/eprint/185026 |
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