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Asian Journal of Animal Sciences

Year: 2016 | Volume: 10 | Issue: 1 | Page No.: 39-48
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Authors

Aris Haryanto

Aris Haryanto

Vera Wati

Suhendra Pakpahan

Nastiti Wijayanti

Keywords

Research Article

Molecular Pathotyping of Newcastle Disease Virus from Naturally Infected Chickens by RT-PCR and RFLP Methods

Aris Haryanto Aris Haryanto's LiveDNA, Vera Wati, Suhendra Pakpahan and Nastiti Wijayanti
Newcastle 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.
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