Article history: Received: 23-10-2016, Accepted: 08-02-2017, Published online: 21-03-2017
Corresponding author: Partha Sarathi Mandal
E-mail: firstname.lastname@example.orgCitation: Mandal PS, Mukhopadhayay SK, Pradhan S, Mondal S, Jana C, Patra NC, Hansda RN (2017) Development of nested polymerase chain reaction-based diagnosis of duck enteritis virus and detection of DNA polymerase gene from non-descriptive duck breeds of West Bengal, India, Veterinary World, 10(3): 336-341.
Aim: The study was undertaken to detect the clinical signs, postmortem lesions of embryonated duck plague (DP) infected eggs, and histopathological changes of chorioallantoic membrane (CAM) in non-descriptive ducks of West Bengal with special reference to standardize nested polymerase chain reaction (PCR).
Materials and Methods: After postmortem of suspected carcasses, samples were collected for virus isolation and identification through specific pathogen free (Khaki Campbell) embryonated duck eggs. PCR was also done as confirmatory test after doing postmortem of duck embryos. DP specific nested PCR was standardized for better confirmation of the disease. Sensitivity of nested primers was also tested for DP virus.
Results: Gross, postmortem and histopathological changes were prominent in dead embryos. First set of primer was able to detect 602 bp fragments of DNA polymerase gene of duck enteritis virus from infected CAM. Subsequently, a DP specific nested PCR which was very much sensitive for very small amount of viral genome was successfully standardized. After NCBI blast nucleotide sequence of nested PCR product (Accession No. HG425076) showed homology with the sequences data available in GenBank.
Conclusion: The study concludes that PCR assay is very much helpful to diagnose DP disease and developed nested PCR is a double confirmatory diagnostic tool for DP.
Keywords: chorioallantoic membrane histopathology, nested polymerase chain reaction, polymerase chain reaction.
1. ICTV. (2015) Virus taxonomy. EC 47, London, UK. Available from: http://www.ictvonline.org/virustaxonomy.asp, Last accessed on 16/10/2016.
2. Mukherji, A., Das, M.S., Ghosh, B.B. and Ganguly, J.L. (1963a, b) Duck plague in West Bengal. Part I & II. Indian Vet. J., 40: 457-462.
3. Durasiwami, J., Rajendran, M.P., John, M., Ganeshmurthy, M. and Rao, C.V.N. (1979) Duck plague in Tamil Nadu. Indian Vet. J., 56: 1-5.
4. Rajan, A., Nair, M.K., Maryamma, K.I. and Valsala, K.V. (1980) Studies on the epidemiology, symptoms and pathoanatomy of duck plague infection (duck virus enteritis). Indian Vet. J., 57: 12-15.
5. Bhowmik, M.K. and Chakraborty, A.K. (1985) Occurrence of duck plague in ducklings of West Bengal. Indian J. Poult. Sci., 20: 115-118.
6. Ahamed, M.M., Hossain, M.T., Rahman, M., Nazir, K.H.M., Khan, M. F.R., Parvej, M.S., Ansari, W.K., Noor-A-Alahi Chiste, M.N., Amin, K.B., Hossen, M.L., Ahmed, S. and Rahman, M.B. (2015) Molecular characterization of duck plague virus isolated from Bangladesh. J. Adv. Vet. Anim. Res., 2(3): 296-303. [Crossref]
7. Shawky, S., Sandhu, T. and Shivaprasad, H.L. (2000) Pathogenicity of a low-virulence duck virus enteritis isolate with apparent immunosuppressive ability. Avian Dis., 44: 590-599. [Crossref] [PubMed]
8. Hanan, A., Ghada, M., Sadek, E., Afan, E.G., Nahed, I.M., Ekram, S.M. and Norhan, N.M. (2014) Preparation and evaluation of combined inactivated duck vaccine against salmonellosis, duck plague and duck hepatitis. Nat. Sci., 12: 142-147.
9. Kumar, N.V., Reddy, Y.N. and Rao, M.V.S. (2004) Enzyme linked immunosorbent assay for detection of duck plague virus. Indian Vet. J., 5: 481-483.
10. Wu, Y., Cheng, A., Wang, M., Zhang, S., Zhu, D., Jia, A.B., Luo, B.Q., Chen, Z. and Chen, X. (2011) Serologic detection of duck enteritis virus using an indirect ELISA based on recombinant UL55 protein. Avian Dis., 55: 626-632. [Crossref] [PubMed]
11. Konch, C., Upadhyaya, T.N., Goswami, S. and Dutta, B. (2009) Studies on the incidence and pathology of naturally occurring duck plague in Assam. Indian J. Vet. Pathol., 33: 213-215.
13. Hanaa, A., Samadony, E., Laila, A., Salama, E.T. and Afaf, A.K. (2013) Molecular characterization of circulating duck viral enteritis in Egypt during 2012-2013. Br. J. Poult. Sci., 2: 38-44.
14. Wang, G., Qu, Y., Wang, F., Hu, D., Liu, L., Li, N., Yue, R., Li, C. and Liu, S. (2013) The comprehensive diagnosis and prevention of duck plague in northwest Shandong province of China. Poult. Sci., 92: 2892-2898. [Crossref] [PubMed]
15. Bancroft, J.D. and Stevens, A. (2013) Theory and Practice of Histological Techniques. 7th ed. Elsevier, Beijing. p80-253.
16. Mondal, B., Rasool, T.J., Ram, H. and Mallanna, S. (2010) Propagation of vaccine strain of duck enteritis virus in a cell line of duck origin as an alternative production system to propagation in embryonated egg. Biologicals, 38: 401-406. [Crossref] [PubMed]
17. Thompson, J.D., Higgins, D.G. and Gibson, T.J. (1994) Clustal-w-improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res., 22(22): 4673-4680. [Crossref]
18. Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. and Kumar, S. (2013) MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol. Biol. Evol., 30(12): 2725-2729. [Crossref] [PubMed] [PMC]
19. Nei, M. and Kumar, S. (2000) Molecular Evolution and Phylogenetics. Oxford University Press, New York. [PMC]
20. Felsenstein, J. (1985) Confidence limits on phylogenies: An approach using the bootstrap. Evolution, 39: 783-791. [Crossref]
21. Saitou, N. and Nei, M. (1987) The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol. Biol. Evol., 4: 406-425. [PubMed]
22. Sarkar, A.J. (1982) Duck plague in Bangladesh: Isolation and identification of the etiological agent. Indian Vet. J., 59: 669-674.
23. Dlugosch, D., Eis-Hubinger, A.M., Kleim, J., Kaiser, R., Bierhoff, E. and Schneweis, K.E. (1991) Diagnosis of acute and latent varicella-zostervirus infections using the polymerase chain reaction. J. Med. Virol., 35: 136-141. [Crossref] [PubMed]
24. Williams, R.A., Bennett, M., Bradbury, J.M., Gaskell, R.M., Jones, R.C. and Jordan, E.T.W. (1992) Demonstration of sites of latency of infectious laryngotracheitis virus using the polymerase chain reaction. J. Gen. Virol., 73: 2415-2420. [Crossref] [PubMed]
25. Grondahl-Yli-Hannuksela, K., Vuononvirta, J., Barkoff, A.M., Viander, M., Van Der Meeren, O., Mertsola, J. and He, Q. (2012) Gene polymorphism in toll like receptor 4: Effect on antibody production and persistence after acellular pertussis vaccination during adolescence. J. Infect. Dis., 205(8): 1214-1219. [Crossref] [PubMed]
26. Priyadharshini, C.V., Parthiban, M., Kumar, K. and Nireesha, G. (2015) Neucleotide sequence and restriction enzyme analysis of field and vaccine strains of duck plague virus in India. Indian J. Comp. Microbiol. Iimmunol. Infect. Dis., 36(1): 14-17.
27. Liu, S., Chen, S., Li, H. and Kong, X. (2007) Molecular characterization of the herpes simplex virus 1 (HSV-1) homologues, UL25 to UL30, in duck enteritis virus (DEV). Gene, 401: 88-96. [Crossref] [PubMed]
28. Xie, W., Cheng, A., Wang, M., Chang, H., Zhu, D. and Luo, Q. (2010) Molecular cloning and characterization of the UL31 gene from duck enteritis virus. Mol. Biol. Rep., 37: 1495-1503. [Crossref] [PubMed]
29. Cao, X., Cheng, A. and Wang, M. (2012) Bioinformatics Analysis of Duck Plague Virus UL41 Gene. Computational and Information Sciences (ICCIS). DOI: 10.1109/ICCIS.2012.93. [Crossref]30. Wu, Y., Cheng, A.C., Wang, M.S., Zhu, D.K. and Chen, X.Y. (2011) Molecular cloning and phylogenetic analysis of the duck enteritis virus UL55 gene. Adv. Mater. Res., 204-210: 663-671. [Crossref]