Identifikasi Epitop Immunodominan Virus Dengue-3 dengan Peptida Sintetik: Identification of the virus dengue-3 epitope's immunodominan using the synthetic peptide
| Main Author: | Perpustakaan UGM, i-lib |
|---|---|
| Format: | Article NonPeerReviewed |
| Terbitan: |
[Yogyakarta] : Universitas Gadjah Mada
, 2002
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| Subjects: | |
| Online Access: |
https://repository.ugm.ac.id/24623/ http://i-lib.ugm.ac.id/jurnal/download.php?dataId=7601 |
Daftar Isi:
- Di negara-negara tropis, infeksi virus dengue merupakan problem yang serius dibidang kesehatan. Identifikasi virus dengue pada penderita dapat dilakukan dengan immunoesai menggunakan peptida sintetik untuk menentukan epitop virus dominan. Penelitian ini diawali dengan mensistesa 161 peptida sintetik dengan metode Osatomi dan Sumiyoshi menggunakan alat Peptide Synthesizer, Auto-Spot Robot ASP 222 (Abimed). Setiap peptida terdiri dan 15 asam amino dengan 12 asam amino yang saling tumpang tindih. Peptida yang terbentuk ditempelkan pada membran nitroselulosa untuk spot immunoassay terhadap masing-masing 6 serum penderita terinfeksi dengue, 6 serum normal, dan 6 serum tidak terinfeksi dengue. Hasilnya menunjukkan bahwa ada 9 peptida yang mempunyai reaksi spesifik terhadap serum penderita terinfeksi, yakni EGLSGATWVDVVLEH (asam amino nomer 13-27), VCKHTYVDRGWGNGC (91-105), SEIGKVVQHENLKYT (124-138), TVHTGDQHQVGNETQ (142-156), TLGLECSPPTGLDFN (178-192), KGEDAPCKIPFSTED (325-339), PFSTEDGQGKAHNGR (334-348), GARRMAILGDTAWDF (406-420) dan KIGIGVLLTWIGLNS (454-468) Atas dasar hasil spot immunoassay kemudian di sintesis 11 peptida dalam bentuk Iarutan yang digunakan untuk ELISA terhadap 15 serum normal, 22 serum penderita non infeksi dan 37 serum penderita terinfeksi. Hasilnya menunjukkan bahwa peptida BTLDDIELQKTEATQLA, BPFSEDGOGKAHNGR dan BKKEEPVNIEAEPPFG memberikan reaksi yang sangat positifterhadap serum terinfeksi. Kata kunci : peptida sintetik, virus dengue, spot immunoassay Keterangan : A = Alanin, C Sistein, D Asam aspartm, E = Asam glutamat, F = Fenilalanin, G = Gliserin, H Histiclin, I = Isoleusin, K = Lisin, L Leusin, N = Asparagin, P = Prulin, Q = Glutamin, R Arginin, S = Serin, T = Treonin, V = Vann, W = Triptofan, Y = Tirosin ABSTRACT Dengue virus infection has been known as an important health problem in many tropical countries, because of increasing number of patients, expansion of epidemic areas and emergence of severe clinical manifestations. Dengue virus consists of 3 structural and 7 nonstructural proteins. The major virion surface protein, the envelope protein, is the most important antigen with regards to virus biology and humoral immunity. Synthetic peptide derived from the envelope protein gene sequence can be used to identify region of the envelope protein that elicits antibodies. One hundred and sixty-one synthetic peptides were synthesized based on sequence of protein envelope dengue-3 virus published by Osatomi and Sumiyoshi. Each peptide consists of 15 amino acids with the last 12 amino acids overlapped. Peptide synthesizer, auto-spot robot ASP 222 (Abimed), was used for synthesis using solid spot method developed by Ronald Frank. Synthetic peptides attached to nitrocellulose membrane were used for spot immunoassay of 6 normal human sera, 6 dengue infected human sera and 6 Ket Words:Peptida sinc,Spos anfsporpreti