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


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.


Newcastle Disease virus; organ sample; amplification

Full Text:



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:


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.


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.


Article Metrics

Abstract views : 5624 | views : 5203


  • There are currently no refbacks.

Copyright (c) 2017 Jurnal Sain Veteriner

Creative Commons License
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


HP. 0895363078367

Jurnal Sain Veteriner is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.


web stats View My Stats