Pathotyping of Local Isolates Newcastle Disease Virus from Field Specimens by RT-PCR and Restriction Endonuclease Analysis

Abstract

Newcastle Disease is one of the serious viral infection disease in poultry, especially chicken. The etiological agent of Newcastle Disease is Newcastle Disease Virus (NDV). The objective of this research was to develop a method for determining pathotype of NDV in various species of poultry from field specimens rapidly and efficiently by using RT-PCR and REA method. This study was conducted for identification, differentiation and pathotyping of NDV from local isolates by using amplification of RT-PCR followed by REA of F protein encoding gene of NDV. To differentiate virulent and avirulent NDV, the RT-PCR product of F gene in size of 767 basepairs (bp) was analyzed by Hin 1l and Bam HI restriction enzymes. RT-PCR amplification for F gene NDV generated DNA fragments in size of 767bp. RT-PCR and REA method by using Hin 1l and Bam HI enzymes can be used to determine pathotype of NDV from field specimens, for total ten field specimen samples of NDV, three samples are avirulent NDV and seven samples are virulent NDV. RT-PCR and REA methods are appropriate with DNA sequencing method to determine the NDV.