DETEKSI VIRUS AVIAN INFLUENZAPADA PETERNAKAN AYAM PETELUR KOMERSIAL YANG MENERAPKAN BEBERAPA PROGRAM VAKSINASI AI DENGAN METODE REVERSETRANSCRIPTION POLYMERASE CHAIN REACTION

Successful rate of avian influenza (AI) vaccination programs is varied. Avian influenza vaccination will prevent morbidity and protect the chicken from infection of high pathogenic avian influenza (HPAI). The aim of this research was to detect AI virus in layer chicken farmthat had applied AI vaccination program by using reverse-transcription polymerase chain reaction (RT-PCR) method. This research used 3 layer farms on different locations which have implemented homolog vaccine killed subtype H5N1 as its vaccination program. Farm A (SL-3) vaccinated at 4, 12, and 18 weeks old chick, Farm B (SL-4) vaccinated at 5 and 15 weeks old chick, and Farm C vaccinated at 5, 15, and 40 weeks old chick. Twenty five chicken samples in each group were taken for cloacal swabs in every 10 weeks, and the observation of the chicken condition was conducted at 18, 28, 38, 48, and 58 weeks old chick. Cloacal swab samples are then extracted to obtain RNA and performed RT-PCR process using RNA extraction kit from Invitrogen with the forward primer (5'-GCACTTGATATTGT GGATTCTTGATC-3') and reverse primer (5'- AGTAGAAACAAGGTAGTTTTTTACTCC-3') which will amplify the M gene with a length of 200bp and a forward primer (5'- GGAATGCCCCAAATATGTGAAATC-3') and reverse primer (5'- TCTACCATTCCCTGCCATCC-3') which will amplify H5 gene with a length of 545bp. The results product was observed using gel electrophoresis with 1.5% concentration. The results showed that vaccination at Farm A (SL-3) could detect M gen at 38 weeks old which the sample numbers were 1,2,3,4,5,18,19,20,21,22, 23,24,25. Farm B detected M gen at 38 weeks old which the sample numberswere 1-25 except sample 10. Farm C detected M gen at 28 weeks old which sample numbers were 1-25. Based on the result, vaccinated chickens remain asymptomatic which indicate AI virus positive, however AI virus can be amplified on 10-20 weeks post-vaccination, on 28 and 38 weeks old to be precis