Asian Journal of Animal Sciences1819-1878xxxx-xxxxxAsian Network for Scientific Information10.3923/ajas.2016.39.48HaryantoAris WatiVera PakpahanSuhendra WijayantiNastiti 12016101Newcastle disease is an infectious disease in poultry caused by the Newcastle Disease Virus (NDV) which belongs to the family Paramyxoviridae. The NDV can infect various species of birds, both domestic and wild birds with highly morbidity and mortality. Based on the pathotype, NDV can be classified into velogenic (high), mesogenic (moderate) and lentogenic (low) pathotypes, however, clinical symptoms appear on infected birds are often similar that it is difficult to distinguish. In this study, molecular pathotyping of NDV has been done to ten virus isolates that isolated from various type of chicken which naturally infected by NDV using a combination method of RT-PCR and RFLP. It was then compared with quantification of Mean Death Time (MDT) value and DNA sequencing which is commonly method used for NDV pathotyping. The aim of this study was to develop a rapid, precise and efficient method for NDV pathotyping by combination method of RT-PCR amplification and RFLP using HinfI restriction enzyme. The results of NDV pathotyping showed that combination of RT-PCR amplification and RFLP using HinfI enzyme can be used well to differentiate the pathotype of virulent NDV versus avirulent NDV rapidly, accurately and efficiently. A total of ten NDV samples isolated from naturally NDV infected chickens showed that three samples are avirulent NDV, while seven other samples are virulent NDV. In compared with other pathotyping methods, namely MDT quantification and DNA sequencing, the molecular NDV pathotyping by RT-PCR and RFLP methods showed similar conclusion and consistent with highly compatibility.]]>Ahmadi, E., M. Ahmadi, S.A. Pourbakhsh and A. Talebi,20132138142Alexander, D.J.,200142522Alexander, D.J.,20032003pp: 63-99pp: 63-99Alexander, D.J. and D.A. Senne,2008Paramyxo virus infections.]]>2008pp: 75-100pp: 75-100Alexander. D.J.,19911991pp: 496 -519pp: 496 -519Beard, C.W. and R.P. Hanson,19841984pp: 452-470pp: 452-470Berg, J.M., J.L. Tymoczko and L. Stryer,20066th Edn.,Pages: 103Pages: 103Haryanto, A., M. Purwaningrum, S, Verawati, S.H. Irianingsih and N. Wijayanti,2015148590Haryanto, A., D. Kristiawan, S.H. Irianingsih and D.W. Yudianingtyas,201314387393, (In Indonesian)Heine, H and L. Trinidad,20062006Kant, A., G. Koch, D.J. Van Roozelaar, F. Balk and A. Ter Huurne,199726837849Keusch, G.T., M. Pappaioanou, M.C. Gonzalez, K.A. Scott and P. Tsai,2009pp: 165-186pp: 165-186Krishnamurthy, S. and S.K. Samal,19987924192424Lamb and D. Kolakofsky,19961996pp: 1177-1203pp: 1177-1203Lee, E.K., W.J. Jeon, J.H. Kwon, C.B. Yang and K.S. Choi,2009134241248Marin, M.C., P. Villegas, J.D. Bennett and B.S. Seal,199640382390Mase, M. and K. Kanehira,20127416611664Nagai, Y.,199318187Nanthakumar, T., R.S. Kataria, A.K. Tiwari, G. Buchaiah and J.M. Kataria,200024275286Naveen, K.A., S.D. Singh, J.M. Kataria, R. Barathidasan and K. Dhama,20134512311236Oberdorfer, A. and O. Werner,199827237243Olav, S., O.S. Deleeuw, G.L. Hantog, N. Ravenshost and B.P. Peeters,20058617591769Romer-Oberdorfer, A., O. Werner, J. Veits, T. Mebatsion and T.C. Mettenleiter,20038431213129Seal, B.S., D.J. King and J.D. Bennett,19953326242630Susta, L., P.J. Miller, C.L. Afonso and C.C. Brown,201148349360Kou, Y.T., L.L. Chueh and C.H. Wang,19996111911195Tirumurugaan, K.G., S. Kapgate, M.K. Vinupriya, K. Vijayarani, K. Kumanan and S. Elankumaran,20112011Wehmann, E., J. Herczeg, A. Ballagi-Pordany and B. Lomniczi,19971514301433Fatchiyah, E.L. Arumingtyas, S. Widyarti and S. Rahayu,2011pp: 46-49pp: 46-49Tabbu, C.R.,2000Volume 1,pp: 164-186pp: 164-186