Newcastle Disease Virus Detection from Chicken Organ Samples Using Reverse Transcriptase Polymerase Chain Reaction
Lehgarubini Shanmuganathan(1*), Dito Anggoro(2), Michael Haryadi Wibowo(3)
(1) Fakultas Kedokteran Hewan, Universitas Gadjah Mada
(2) Fakultas Kedokteran Hewan, Universitas Gadjah Mada
(3) Fakultas Kedokteran Hewan, Universitas Gadjah Mada
(*) Corresponding Author
Abstract
Newcastle disease (ND) is a systemic, viral respiratory disease that is acute and easily transmitted which affects various types of poultry, especially chickens. Diagnosis of ND which generally involves virus isolation and subsequent identification with serological assays has limitations that needs more time. This research was aimed to detect Newcastle Disease virus (NDV) in chickens suspected with ND using the Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) technique. Nine chicken organ samples such as lien, trachea, and lungs were collected from chicken farms diagnosed with ND. The organ samples were processed and the targeted viral RNA was extracted using the RNA extraction kit. Genome amplification was performed with RT-PCR using specific
primers to target the F gene. Amplification results produced an amplicon product of 565 base pairs (bp). PCR product samples were then visualised using agar gel electrophoresis and viewed using the unified gel documentation system. Amplification results show nine samples positive for the DNA bands corresponding to the targeted band of the NDV F gene fragment. The results of this research confirm that the RT-PCR method is applicable for NDV detection from chicken organ samples.
Keywords
Full Text:
PDFReferences
Alexander, D.J. 2000. Newcastle disease and other
avian paramyxoviruses. Revue Scientifique
et Technique de L`Office International Des
Epizooties 19(2): 443-462.
Alexander, D. Newcastle disease, other avian
paramyxoviruses, and pneumovirus infections.
In: Diseases of Poultry. Saif, Y.M., Barnes,
H.J., Glisson, J.R., Fadly, A.M., McDougald,
L.R., Swayne, D., (eds)., Iowa State University
Press, Ames. 63–99.
Alexander, D.J., Bell, J.G. and Alders, R.G. 2004. A
Technology Review: Newcastle Disease. FAO
Animal Health and Production Paper, 161:1-15.
Amanu, S. and Rohi, O.K. 2005. Studi Serologis
dengan Uji Hambatan Hemaglutinasi terhadap
Angsa yang dapat bertindak sebagai pembawa
Newcastle Disease di D.I.Yogyakarta. Jurnal
Sain Veteriner, 1: 8-12.
Angeliya, L. 2014. Analisis Phylogenic Tree Dan
Konsensus Sequence Virus Newcastle Disease
Isolat Asal Wilayah Kerja Balai Veteriner
Lampung Tahun 2010-2013. In: Prosiding
Seminar Nasional Sains dan Inovasi Teknologi
Pertanian, Balai Pengkajian Teknologi
Pertanian (BPTP), Lampung. 474-481.
Cattoli, G., Susta, L., Terregino, C., and Brown, C.
Newcastle disease: a review of field
recognition and current methods of laboratory
detection. Journal of Veterinary Diagnostic
Investigation, 23(4): 637–656.
de Leeuw, O.S., Koch, G., Hartog, L., Ravenshorst,
N. and Peeters, B.P. H. 2005. Virulence of
Newcastle disease virus is determined by the
cleavage site of the fusion protein and by
both the stem region and globular head of
the haemagglutinin–neuraminidase protein.
Journal of General Virology, 86: 1759–1769.
Dharmayanti, N.L.P.I., Hartawan, R., Hewajuli, D.A.
and Indriani, R. 2014. Phylogenetic Analysis
of Genotype VII of Newcastle Disease Virus
in Indonesia. African Journal of Microbiology
Research, 8(13): 1368-1374
Gohm, D.S., Thür, B. and Hofmann, M. A. 2000.
Detection of Newcastle disease virus in organs
and faeces of experimentally infected chickens
using RT-PCR. Journal of Avian Pathology,
(2): 143-152
Haque, M.H., Hossain, M. T., Islam, M. T., Zinnah,
M. A., Khan, M. S. R. and Islam, M.A. 2010.
Isolation and Detection of Newcastle Disease
Virus From Field Outbreaks In Broiler And
Layer Chickens By Reverse Transcription–
Polymerase Chain Reaction. Bangladesh
Journal of Veterinary Medicine, 8(2): 87–92.
Kant, A., Koch, G., Van Roozelaar, D.J., Balk, F.
and Ter Huurne, A. 1997. Differentiation of
virulent and non‐virulent strains of Newcastle
disease virus within 24 hours by polymerase
chain reaction. Avian Pathology Journal,
(4): 837-849
Kencana, G.A.Y., Kardena, I.M., and Mahardika,
I.G.N.K. 2012. Diagnosis Confirmation of
Newcastle Disease on Native Chicken in Bali
Using RT-PCR Method. Jurnal Kedokteran
Hewan, 6(1): 28-31
Kho, C.L., Tan, W.S., Tey, B.T. and Yusoff, K.
Newcastle Disease Virus Nucleocapsid
Protein. Journal of General Virology, 84:
–2168.
Kryger, K.N., Thomsen, K.A., Whyte, M.A. and
Dissing, M. 2010. Smallholder Poultry
Production. FAO Smallholder Poultry
Production Paper. 4: 7-25.
MacLachlan, N.J. and Dubovi, E.J. (eds). 2010.
Fenner’s Veterinary Virology. 4th Edition.,
Academic Press Elsevier, London. 300, 304-
, 311-314.
Miller, P.J., Decanini, E.L. and Afonso, C.L. 2010.
Newcastle disease. Infection, Genetics and
Evolution, 10(1): 26–35.
Oberdorfer, A. and Werner, O. 1998. Newcastle
disease virus: Detection and characterization
by PCR of recent German isolates differing
in pathogenicity. Journal of Avian Pathology,
(3): 237-243.
Office International des Epizooties (OIE). 1996.
Manual of Standards for Diagnostic Tests and
Vaccines. 3rd Edition., OIE Manual, 161-165.
Office International des Epizooties (OIE). 2008.
Manual of Diagnostic Tests and Vaccines
for Terrestrial Animals. 6th Edition., OIE
Terrestrial Manual, 1: 576-582.
Pelt-Verkuil, E.V., Belkum, A.V., and Hays, J.P.
Principles and Technical Aspects of PCR
Amplification. Springer Science & Business
Media. 20.
Rabalski, L., Smietanka, K., Minta, Z., and Szewczyk,
B. 2014. Detection of Newcastle Disease Virus
Minor Genetic Variants by Modified Single-
Stranded Conformational Polymorphism
Analysis. BioMed Research International,
(632347): 1-8.
Rio, D. C., Ares, M., Hannon, G.J. and Nilsen, T.W.
RNA: A Laboratory Manual. Cold Spring
Harbour Laboratory Press. New York. 17.
Saif, Y.M. 2011. Diseases of Poultry. 12th Edition.,
Iowa, John-Wiley and Sons Publishing Inc.
-93.
Shirvan, A.S. and Mardani, K. 2014. Molecular
Detection of Infectious Bronchitis and
Newcastle Disease Viruses in Broiler Chickens
with Respiratory Signs using Duplex RT-PCR.
Veterinary Research Forum, 5(4): 319 – 323
Singh, K., Jindal, N., Gupta, S.L., Gupta, A.K.
and Mittal D. 2005. Detection of Newcastle
Disease Virus Genome from the Field Outbreaks in Poultry by Reverse Transcription– Polymerase Chain Reaction. International
Journal of Poultry Science, 4(7): 472-475.
Tabbu, C.R. 2000. Penyakit Ayam dan
Penanggulangannya: Volume 1. Penerbit PT
Kanisius, Yogyakarta, 49-51.
Wibowo, M.H. and Amanu, S. 2010. Perbandingan
beberapa Program Vaksinasi Penyakit
Newcastle pada Ayam Buras. Jurnal Sain
Veteriner, 28(1): 27-35.
Wibowo, M.H., Untari, T. and Wahyuni, A.E.T.H.
Isolation, Identification, Physical, And
Biological Character Of Newcastle Disease
Virus Isolated From Field Cases. Jurnal Sain
Veteriner, 13(4): 425-433.
DOI: https://doi.org/10.22146/jsv.29300
Article Metrics
Abstract views : 6313 | views : 5562Refbacks
- There are currently no refbacks.
Copyright (c) 2017 Jurnal Sain Veteriner
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Jurnal Sain Veteriner Indexed by
Copyright of JSV (Jurnal Sain Veteriner) ISSN 0126-0421 (print), ISSN 2407-3733 (online).
Fakultas Kedokteran Hewan, Universitas Gadjah Mada
Jl. Fauna No.2, Karangmalang, Yogyakarta
Phone: 0274-560862
Fax: 0274-560861
Email: jsv_fkh@ugm.ac.id
HP. 0895363078367
Jurnal Sain Veteriner is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
View My Stats