Research Article | OPEN ACCESS
Differentiation between Vaccinal and Iranian Virulent Isolates of Newcastle Disease Virus based on F Region Genotyping by HRM Analysis
1Shahdad Dibazar, 1Nariman Sheikhi, 2Farhid Hemmatzadeh, 1Saeed Charkhkar and 3Seyed Ali Pourbakhsh
1Department of Clinical Science, College of Veterinary, Tehran Science and Research Branch, Islamic Azad University, Tehran, Iran
2School of Animal and Veterinary Sciences, the University Adelaide, South Australia, Australia
3Department of Poultry Diseases Research, Razi Vaccine and Serum Research Institute, Karaj, Iran
International Journal of Animal and Veterinary Advances 2014 6:150-155
Received: March 13, 2014 | Accepted: April 11, 2014 | Published: December 20, 2014
Abstract
The purpose of this study is investigate Differentiation Between Vaccinal and Iranian Virulent Isolates of Newcastle Disease Virus based on F Region Genotyping by HRM Analysis. Discrimination of circulating virulent strains of Newcastle Disease Virus (NDV) from low pathogenic and vaccine stains is the basis for implementation of strategies to control and eradication of that aims at the eradication of NDV in poultry. At the present study the applicability of Real time RT-PCR followed High-Resolution Melting-Curve Analysis (HRM) was evaluated for differentiation of pathogenic ND viruses from the live vaccinal strains. Five virulent NDV isolates and 6 live vaccines samples of NDV were used to develop and evaluate the tests. Based on the nucleotide sequence of Fusion (F) gene, 3 primer pairs (B, E and H) were designed and utilized for HRM analysis. The melting temperature of vaccinal strains higher than the virulent isolates were obtained using the B and H primer pairs, but it was not the same as E. Based on the analysis of HRM curves in this study, the B and H primer pairs could better differentiate the vaccinal strains from each other and from virulent isolate than E. As it is found in this study, HRM analysis can differentiate virulent NDVs from live vaccines. In comparison to the existing methods for discrimination of different pathotypes of the ND viruses, the developed test takes a shorter time, needs less infrastructure and facilities and costs significantly less than the other traditional methods like intra cerebral pathogenicity index (ICPI).
Keywords:
Differentiation, HRM Analysis, Newcastle disease virus, primer, vaccinal virus, wild virus,
Competing interests
The authors have no competing interests.
Open Access Policy
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
Copyright
The authors have no competing interests.
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ISSN (Online): 2041-2908
ISSN (Print): 2041-2894 |
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