387 Amplifikasi Gen Penyandi Protein Fusion Virus Tetelo dari Spesimen Lapangan dengan Metode OneStep RT-PCR. (AMPIFICATION OF FUSION PROTEIN ENCODING GENE OF NEWCASTLE DISEASE VIRUS FROM FIELD SPECIMENS BY ONESTEP RT-PCR METHOD) Aris Haryanto 1 , David Kristiawan 1 , Sri Handayani Irianingsih 2 , Dini Wahyu Yudianingtyas 3 1 Bagian Biokimia, Fakultas Kedokteran Hewan, Universitas Gadjah Mada, jln. Fauna 2, Karangmalang, Yogyakarta 55281. Telp. 0274-560865, Email: arisharyanto@yahoo.com 2 Laboratorium Virologi dan Serologi, Balai Besar Veteriner Wates, Yogyakarta. 3 Laboratorium Virologi dan Bioteknologi, Balai Besar Veteriner Maros, Sulawesi Selatan. ABSTRAK Virus tetelo atau Newcastle Disease (ND) termasuk dalam famili Paramyxoviridae yang mempunyai genom RNA beruntai tunggal dan merupakan virus beramplop. Pada amplop virus tetelo terdapat dua protein utama, yaitu protein Haemagglutinin/ Neuraminidase (H/N) dan protein Fusion (F). Metode diagnosis molekuler berbasis OneStep RT-PCR merupakan metode diagnosis yang umum digunakan untuk mendiagnosis penyakit tetelo pada unggas. Pada penelitian ini diagnosis penyakit tetelo dilakukan dengan mengamplifikasi gen penyandi protein F virus tetelo langsung dari spesimen lapangan tanpa melalui proses inokulasi dan propagasi virus tetelo pada telur ayam berembrio (TAB). Tujuan dari penelitian ini adalah melakukan diagnosa cepat tetelo langsung dari spesimen lapangan berdasarkan hasil amplifikasi gen penyandi protein F dengan metode OneStep RT-PCR, sehingga dari hasil penelitian ini diharapkan dapat digunakan untuk membantu proses identifikasi virus tetelo langsung dari spesimen lapangan. Hasil penelitian menunjukkan bahwa dari 15 sampel virus yang isolasi dari swab trachea unggas yang secara klinis menunjukkan gejala terinfeksi virus tetelo sebanyak 12 sampel atau 80% dari sampel uji terbukti positif terinfeksi virus tetelo yang ditunjukkan dengan fragmen DNA hasil amplifikasi sebesar 362 bp. Metode OneStep RT-PCR merupakan metode diagnosis yang cepat dan efektif dalam mendiagnosis penyakit tetelo menggunakan spesimen lapangan. Kata kunci: virus tetelo, gen F, spesimen lapangan, RT-PCR. ABSTRACT Tetelo or Newcastle Disease (ND) virus is belong to the family Paramyxoviridae, which has a single stranded RNA (ss RNA) genome and it has viral envelope. The viral envelope consists of two major proteins, namely Haemagglutinin/Neuraminidase (H/N) and Fusion (F) protein. Molecular diagnostic methods OneStep RT-PCR is a commonly diagnostic tool that used to diagnose of Tetelo in poultry. In this study the diagnosis of Tetelo is accomplished by amplification of F protein encoding gene of Tetelo virus directly from field specimens without inoculation and propagation of Tetelo virus into embryonated chicken eggs. The objective of this study is to conduct rapid diagnosis of Tetelo virus directly from the field specimens based on the amplification of the F gene by a OneStep RT-PCR method, so that the results of this study can be used to assist in the identification of Tetelo virus directly from the field specimens. The results showed that from the 15 samples of virus which isolated from tracheal swabs of clinical poultry showing symptoms of Tetelo virus infection, a total of 12 samples from 15 tested sampels (80%) are positive tested for Tetelo virus infection. They were indicated by the amplification product of DNA fragments in size of 362 bp. OneStep RT-PCR is a method for rapid and effective diagnosis which can be used to diagnose of Tetelo virus directly from field specimens. Keywords: Tetelo virus, F gene, field specimen, RT-PCR. Jurnal Veteriner September 2013 Vol. 14 No. 3: 387-393 ISSN : 1411 - 8327