156 Deteksi Molekuler Virus Infectious Bursal Disease (IBD) pada Samp l Bursa Fabrisius yang Diperoleh dari Ayam Terdiagnosa Penyakit IBD Molecular Detection of Infectious Bursal Disesase Virus at Bursa Fabrisius Samples Obtained from Chicken Suspected to IBD Infection 1 2 3 1 1 Michael Haryadi Wibowo , Radhiyan Fadiar , Dito Anggoro , Sidna Artanto , Surya Amanu , 1 Agnesia Endang Tri Hastuti Wahyuni 1 2 3 Departemen Mikrobiologi, Mahasiswa Kedokteran Hewan, Mahasiswa Program Studi Sain Veteriner, Fakultas Kedokteran Hewan, UGM. Jln. Fauna No2. Karangmalang, Yogyakarta. Email : Abstract Infectious Bursal Disease (IBD) still existing in the commercial layer and broiler farm in Indonesia. Diagnosis of IBD was done according to specific lesion finding and in ovo inoculation based on macroscopic examination of chicken embrio and further identification using agar gel precipitation test (AGPT). The aim of this research was to applied molecular diagnostic test using reverse-transcriptation polymerase chain reaction (RT-PCR) to confirm IBD virus from BF samples suspected to IBD virus infection. Virus detection using AGPT with chorioallantoic membrane (CAM) and embryo as source of antigen, to choose the best antigen source for AGPT. Five samples of bursa fabricius were collected from commercial farms in Yogyakarta special province which were further processed for virus detection. Confirmation test was performed by RT-PCR method using specific primers targeted to VP2 gene fragment. Positive RT-PCR were subjected to virus isolation on chicken embryonated egg 11-days old, and antibody negative to IBD virus. Inoculation materials were injected at chorioallantoic membrane of chicken embryonated eggs, incubated, and collected from refrigerator at 5 days post infection. Membrane chorioallantois and embryo were harvested and further processed for AGPT. Confirmation test using RT-PCR method that amplified VP2 gene fragment showed that 3 samples out from 5 samples were positive. Serologic assay of AGPT using CAM obtained from positive PCR as source of antigen indicated 2 positive out of 3 samples, meanwhile AGPT using embryo as sources antigen were negative. Based on the research could be concluded that RT-PCR method could be used to detect IBD virus genom from directly BF samples. AGPT test was performed with antigen obtained from CAM indicated better result compared to that embrio. Key words: bursa fabrisius, chorioallantoic membrane, precipitation, amplification, IBD virus. e mhwibowo@ugm.ac.id JURNAL SAIN VETERINER ISSN : 0126 - 0421 JS 33 (2), Desember 2015 V