Pendekatan Diagnosis Avian Influenza Virus dan Newcastle Disease Virus pada Kasus Lapangan Ayam Petelur: Imunopatologis Streptavidin Biotin

Maya Ekaningtias(1*), Hastari Wuryastuty(2), Wasito Wasito(3)

(1) Program Studi Bioteknologi, Sekolah Pasca Sarjana, Universitas Gadjah Mada
(2) Fakultas Kedokteran Hewan, Universitas Gadjah Mada
(3) Fakultas Kedokteran Hewan, Universitas Gadjah Mada
(*) Corresponding Author


Poultry can be infected not only by one type of virus, but also by more than one type of virus. Avian influenza virus (AIV) and Newcastle disease virus (NDV) are two important pathogens in poultry that can cause recurrent outbreaks and often lead to clinical symptoms and pathological lesions similar in poultry with high morbidity and mortality causing significant economic losses to the poultry industry. This study aims to detect and differentiate pathogenic viruses in commercial laying chickens with similar clinical symptoms, especially torticolis and curled toe paralysis, and anatomic pathological lesions, such as foci necrotic hemorrhages in the digestive tract by applying streptavidin-biotin immunohistochemical immunological test (IHC SB). Samples (chickens) were taken from the cases of the disease in poultry in several commercial poultry farms. The layer chickens in which showed clinical symptoms of torticolis and curled toe paralysis, and hemorrhagic lesions in the lungs and / or digestive system are suspected of having AIV and NDV infections. After being necropsied, then lungs and gastrointestinal tract were tested IHC SB. The IHC SB results were examined with a digital microscope camera system. The results of the present study indicated that AIV antigen was detected in the lungs, but not in the digestive tract, whereas, NDV was detected in the digestive tract, but not in the lungs. It was concluded, that the IHC SB can be applied for confirmation of the diagnosis of AIV and NDV and indicated, that the commercial laying chickens with clinical symptoms of torticolis and curled toe paralysis, as well as pathological hemorrhagic lesions in the gastrointestinal tract could be infected by both viruses.


AIV; NDV; IHC SB; torticolis; paralysis

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Anonimus, 2007, Introduction to Antibodies,

(Diakses tanggal 3 februari 2015).

Achenbach, J.E. and Bowen, R.A., 2011, Transmission

of Avian Influenza A Viruses Among Species

in an Artificial Barnyard, PLoS One, 6 (3):


Adi, A.A.A.M., Kardena, I.M, Astawa, N.M. and

Matsumoto, Y., 2012, Pelacakan Secara

Imunohistokimiawi Antigen Virus pada Ayam

yang Diinfeksi dengan Virus Penyakit Tetelo,

J.Vet., 13. (3): 278-283.

Alexander, D.J. and Stuart, J.C., 1982, Isolation of

An Influenza A Virus from Domestic Fowl in

Great Britain, Vet. Rec.,111:416.

Alexander, D. J., 2000, Newcastle Disease and Other

Avian Paramyxoviruses, Rev. Sci. Off.Int.

Epiz., 19, 443-462.

Alexander, D.J. and Senne, D.A., 2008, Newcastle

Disease, Other Avian Paramyxovirus and

Pneumovirus Infection In: Alexander and

Senne, Y.M. (ed.), Disease of Poultry.

Blackwell Publishing. Iowa. 75-92.

Capua, I. and Alexander, D.J., 2009, Ecology,

Epidemiology and Human Health, Implications

of Avian Influenza Virus Infections, Avian Influenza and Newcastle Disease, @Springer-Verlag Italia.

Cattoli, G., Susta, L., Terregino, C. and Brown,

C., 2011, Newcastle Disease : A Review of

Field Recognition and Current Methods of

Laboratory Detection, J.Vet. Diagn. Invest.,

: 637-658.

Chamnanpood , C., Sanguansermsri, D., Pongcharoen,

S. and Sanguansermsri, P., 2011, Detection of

Distribution of Avian Influenza H5N1 Virus

By Immunohistochemistry, Chromogenic

In Situ Hybridization and Real-Time PCR

Techniques In Experimentally Infected

Chickens, Southeast Asian J. Trop. Med. Pub.

Health, 42.(2): 303-310.

Costa-Hurtado, M., Afonso, C.L., Patti J Miller,

Spackman, E., Kapczynski, D.R., Swayne,

D.E., Shepherd, E., Smith, D., Zsak, A. and

Pantin-Jackwood, M., 2014, Virus Interference

between H7N2 Low Pathogenic Avian

Influenza Virus and Lentogenic Newcastle

Disease Virus in Experimental Co-Infections

in Chickens and Turkeys, Vet. Res., 45: 1.

Damayanti, R., Dharmayanti, N.L.P.I, Indriani, R.,

Wiyono, A., Darminto, 2004, Gambaran Klinis

dan Patologis pada Ayam yang Terserang

Flu Burung Sangat Patogenik (HPAI) di

Peternakan di Jawa Timur dan Jawa Barat,

JITV, 9. (2): 128-135.

Farmilo, A.J. and Stead, R.H., 2009, Fixation

and Proccesing. In Educational Guide

Immunohistochemistry Staining Methods, 5th

edition, Dako North America, Carpinteria,


Ge, J., Deng, G., Wen, Z., Tian, G., Wang, Y., Shi,

J., Wang, X., Li, y., Hu, S., 2007, Newcastle

Disease Virus Based Live Attenuated Vaccine

Completely Protecs Chickens and Mice

From Lethal Challenge of Homologous and

Heterologous H5N1 Avian Influenza Viruses,

J. Virol., 150-158.

Gelb, J., Ladman, B.S., Licata, M.J., Shapiro,

M.H., Campion, L.R., 2007, Evaluating

Viral Interference Between Infectious

Bronchitis Virus and Newcastle Disease Virus

Vaccine Strains Using Quantitative Reverse

Transcription-Polymerase Chain Reaction,

Avian Dis., 51:924–934.

Gelb, J., Ladman, B.S., Licata, M.J., Shapiro,

M.H., Campion, L.R., 2007, Evaluating

Viral Interference Between Infectious

Bronchitis Virus and Newcastle Disease Virus

Vaccine Strains Using Quantitative Reverse

Transcription-Polymerase Chain Reaction,

Avian Dis., 51:924–934.

Gu, J,, Xie, Z,, Gao, Z., 2007, H5N1 Infection of The

Respiratory Tract and Beyond: A Molecular

Pathology Study, Lancet, 370: 1137–1145.

Kalthoff, D., Breithaupt, A., Teifke, J.P., 2008, Highly

Pathogenic Avian Influenza Virus (H5N1) in

Experimentally Infected Adult Mute Swans,

Emerg. Infect. Dis., 14: 1267-70.

Kapczynski, D.R., Afonso, C.L. and Miller, P.J.,

, Immune Responses of Poultry to

Newcastle Disease Virus, Develop. Comp.

Immuno., 41:447–453.

Oldoni, I., Brown, C.C., King, D.J., and

other authors, 2005, The Use of In Situ

Hybridization and Immunohistochemistry to

Study The Pathogenesis of Various Newcastle

Disease Virus Strains and Recombinants in

Embryonated Chicken Eggs, Micro. Pathog.,

: 69-75.

Perkins, L.E.L. and Swayne, D.E., 2001, Pathobiology

of A/Chicken/Hong Kong/220/97 (H5N1)

Avian Influenza Virus in Seven Gallinaceous

Species, Vet. Pathol., 38:149-164.

Pattison, M., McMullin, P.F., Bradbury, J.M. and

Alexander, D.J., 2008, Poultry Diseases, 6th

Edition, Saunders Elsevier, China, pp: 298-301.

Post, J., de Geus, D.G., Vervelde, L., Cornelissen,

J.B.W.J. and Rebel, J.M.J., 2013, Systemic

Distribution of Different Low Pathogenic

Avian Influenza (LPAI) Viruses in Chicken, J.

Virol., 10:23.

Quinn, P.J., Markey, B.K., Carter, M.E., Donnelly,

W.J.C. and Leonard, F.C., 2002, Veterinary

Microbiology and Microbial Disease, Blackwell

Science. Iowa. 283-289, 308, 381-387.

Rimmelzwaan, G.F., Kuiken, T., Amerongen, G.V.,

Bestebroer, T.M., Fouchier, R.A.M., Osterhaus,

A.D.M.E., 2001, Pathogenesis of Influenza A

(H5N1) Virus Infection in A Primate Model, J.

Virol, 75: 6687-91.

Ronohardjo, P., Hardjosworo, S., Partoatmojo, S. and

Partadireja, M., 1985, The Identification and

Distribution of Influenza A Virus in Indonesia,

Penyakit Hewan XVII, Semester I: 249-257.

Spackman, E., 2008, A Brief Introduction to The Avian

Influenza Virus, Methods Mol. Biol., 436, 1–6.

Swayne, D.E. and Pantin-Jackwood, M., 2008,

Pathobiology of Avian Influenza Virus

Infections in Birds and Mamals, Blackwell

Publishing Professional 2121 State Avenue,

Ames Iowa 50014, USA: 87-122.

Vascellari, M., Granato, A., Trevisan, L., Basilicata,

L., Toffan, A., Milani, A. and Mutinelli, F., 2007,

Pathologic Findings of Highly Pathogenic

Avian Influenza Virus A/Duck/Vietnam/12/05

(H5N1) on Experimentally Infected Pekin

Ducks, Based on Immunohistochemistry and

In Situ Hybridization, Vet. Pathol., 44: 635-642.

Wakamatsu, N., King, D.J., Kapczynski, D.R.,

Seal, B.S., Brown, C.C., 2006, Experimental

Pathogenesis For Chickens, Turkeys, And

Pigeons of Exotic Newcastle Disease Virus

from An Outbreak in California During 2002–

, Vet. Pathol., 43:925–933.

Wakamatsu, N., King, D.J., Seal, B.S., Brown,

C.C., 2007, Detection of Newcastle Disease

Virus RNA by Reverse Transcription-Polymerase

Chain Reaction Using Formalin-Fixed,

Paraffin-Embedded Tissue and Comparison

with Immunohistochemistry and In Situ

Hybridization, J. Vet .Diagn. Invest., 19:396-400.

Warnke, and Levy, R., 1980, Detection of T and B

Cell Antigens with Hybridoma Monoclonal

Antibodies: Biotin Avidin Horseradish

Peroxidase Method, J. Histochem. Cytochem.,

: 771.

Wasito, R., 1991, Penggunaan Imunositokimia

untuk Diagnosis Penyakit Infeksi, Kursus

Singkat Imunositokimia di PAU Bioteknologi,

Universitas Gadjah Mada, Yogyakarta.

Wasito, R., Wuryastuty, H., Tjahyowati, G., Irianingsih,

S. H., Tyasasmaya, T. and Maes, R. K., 2014,

Detection and Differentiation of Pathogenic

H5 and H7 Influenza A Virus Subtypes in

Indonesian Poultry by Multiplex Reverse

Transcription-Polymerase Chain Reaction,

Biochem. and Biotech. Res., 2(2): 27-31.


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