Evaluation specific primer design for HN and M NDV genes based on Iraqi virulent isolates by used real-time RT-florescent PCR technique

Abstract EN

Background and Aims :Newcastle disease is considered the most contagious poultry disease and may cause severe economic loss in the poultry industry.

The virus belongs to the Avulavirus genus within the family Paramyxoviridae, subfamily Paramyxovirinae, of the order Mononegavirales and is designated avian paramyxovirus-1.

So, Early detection of the virus by used sensitive and specific primer design for detect local outbreak isolate can prevent the spread of disease and avoid huge economic losses.

Towards this goal, in this research, we developed reliable specific primers to reproduce specific amplification for NDV HN and M genes in One-step Real-time RT-Florescent PCR method.

Materials and Methods: in this experimental study , Tow NDV isolate kindly provided by Iraqi center for cancer and medical genetic research , respective RNA was extracted from virus by using viral RNA/DNA extracted kit .

the specific primers were designed according to NCBI data base for NDV HN and M gene sequence aligned by using ApE program and IDT company software Intercalating primer design website and sudsequently to amplify and evaluate by Real-time RT-Florescent PCR one step intercalaing dye .

Results: the fourescent signle and the melting point analysis for amplification the NDV HN and M genes give positive results by specific primer design Conclusion: This study showed that the primer design for NDV HN and M genes are sensitive , specific and accurate by used Real-time RT-Florescent PCR Technique molecular assay for rapid diagnosis of local NDV outbreak isolate .