Zahra Talebi 2, Mohammad mehdi Kiaee Daronkola2, Taregh Eslami Jouybari 2, Abbas Alamian 1*
- Razi Vaccine and Serum Research Institute, Karaj, Iran
- Molecular Department, Ariana Imen Padtan Alborz (AIPA) Biotechnology Company, Tehran, Iran
Email: abbasalemian@yahoo.com
Objectives: Newcastle disease (ND) is caused by a highly contagious virus belonged to the avian paramyxovirus type 1. Different species of birds are affected by the virus; however, distribution of this disease in poultry is considered as a the most important challenge in the industrial farms[1].
The disease appears in three main forms consisted of lentogenic or mild, mesogenic or moderate, and velogenic or very virulent infections. Therefore, symptoms of ND range from mild respiratory symptoms to severe neurological manifestations, which lead to high mortality rates[2]. Scheduled vaccination programs are recommended to control this disease, which may result in misdiagnosis of true infection from vaccine isolates in a farm[3]. Therefore, fabrication of an accurate and rapid test, capable to distinguish pathogenic strain from vaccine strains is crucial for clinicians[4, 5].
This study aimed to design a conventional PCR technique to detect Newcastle virus and distinguish vaccinal strains from virulent strains in poultry farms.
Materials & Methods: Primers were designed based on the conserved region of the “F” gene to amplify a 275-bp fragment for almost all paramyxovirus 1. In addition, specific primers, targeting a 255-bp identical fragment of the “F” gene, were designed to recognize vaccine strains including V4, LaSota, B1, and probably I2. The virus samples were collected from a poultry farm in Mazandaran province. In addition, available vaccines: V4, LaSota, B1, and I2 were included in the study. Total RNA was extracted and the identical fragments were amplified by specific primers after complementary DNA synthesis.
Results and Conclusion: The results of conventional PCR for the paramyxovirus 1-specific showed amplification of a 275-bp fragment for all isolates together with the vaccine strains. In addition, amplification using by the vaccine-strain specific primers showed the presence of a 255-bp fragment of the “F” gene in vaccine strains. The findings confirmed the lack of amplifications of the virulent strains by vaccine-specific primers.
The findings showed that the present molecular setting was able to detect paramyxovirus 1 in both vaccine and wild isolates. Furthermore, the current primers distinguished vaccine strains V4, LaSota, B1, and I2 from virulent strains collected from poultry farm.
Keywords: Poultry; Newcastle disease; Vaccine strain; PCR; Industrial farm
References
- Ganar, K., et al., Newcastle disease virus: Current status and our understanding. Virus Research, 2014. 184: p. 71-81.
- Zhang, D., Z. Ding, and X. Xu Pathologic Mechanisms of the Newcastle Disease Virus. Viruses, 2023. 15, DOI: 10.3390/v15040864.
- Dimitrov, K.M., et al., Newcastle disease vaccines—A solved problem or a continuous challenge? Veterinary Microbiology, 2017. 206: p. 126-136.
- Gallili, G.E. and D. Ben-Nathan, Newcastle disease vaccines. Biotechnology Advances, 1998. 16(2): p. 343-366.
- Nidzworski, D., L. Rabalski, and B. Gromadzka, Detection and differentiation of virulent and avirulent strains of Newcastle disease virus by real-time PCR. Journal of Virological Methods, 2011. 173(1): p. 144-149.
نظرات کاربران