Deteksi Molekuler Avian Influenza Virus dan Newcastle Disease Virus pada Ayam Petelur Komersial dengan Gejala Neurologis Menggunakan sRT-PCR
DOI:
https://doi.org/10.71024/bioindikator.2025.v2i2.377Keywords:
Avian influenza virus, Newcastle disease virus, sRT-PCR, molecular diagnosis, layer chickensAbstract
Viral infections in commercial poultry are not always caused by a single pathogen and may involve concurrent infections by multiple viruses. Avian influenza virus (AIV) and Newcastle disease virus (NDV) are among the most important pathogens affecting poultry health, as they often induce overlapping clinical manifestations and tissue alterations, leading to high mortality rates and substantial economic losses. This study investigated the presence of these viruses in commercial layer chickens exhibiting neurological signs, including torticollis and leg paralysis, accompanied by hemorrhagic lesions in the digestive tract, using a molecular diagnostic approach. Serum and digestive tract samples were collected from affected layer chickens originating from commercial farms with reported disease outbreaks. All samples were analyzed using simplex reverse transcriptase polymerase chain reaction (sRT-PCR). RNA extraction for AIV was performed using a commercial kit, while NDV RNA was isolated using Trizol reagent. Viral detection was carried out using specific primers targeting the nucleoprotein gene of AIV and the conserved region of NDV, producing amplicons of 552 bp and 320 bp, respectively. Amplified products were visualized by electrophoresis on 1.5% agarose gel. The results demonstrated the presence of AIV-specific amplicons in serum samples and NDV-specific amplicons in digestive tract tissues, indicating concurrent infection by both viruses in the examined layer chickens. These findings confirm that sRT-PCR is a reliable molecular tool for the confirmation and differentiation of AIV and NDV infections in poultry cases presenting similar clinical signs and pathological lesions.
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References
Achenbach, J. E., & Bowen, R. A. (2011). Transmission of avian influenza A viruses among species in an artificial barnyard. PLoS ONE, 6(3), e17643.
Alexander, D. J. (2000). Newcastle disease and other avian paramyxoviruses. Revue Scientifique et Technique (OIE), 19, 443–462.
Alexander, D. J., & Senne, D. A. (2008). Newcastle disease and other avian paramyxoviruses. In Diseases of Poultry (12th ed.). Blackwell Publishing.
Alexander, D. J., & Stuart, J. C. (1982). Isolation of an influenza A virus from domestic fowl in Great Britain. Veterinary Record, 111, 416.
Andriyani, N. R., Shanti, A. R., Angeliya, L., Anggita, M., Untari, T., Wahyuni, A. E. T. H., Asmara, W., & Wibowo, M. H. (2022). Deteksi Virus Newcastle Disease pada Burung Merpati (Columba livia) dan Burung Tekukur (Streptopilia chinensis) yang Menunjukkan Gejala Syaraf. Acta Veterinaria Indonesiana, 10(3), 220–227.
Bosco-Lauth, A., Rodriguez, A., Maison, R. M., Porter, S. M., & Root, J. J. (2023). H7N9 influenza A virus transmission in a multispecies barnyard model. Virology, 582, 100–105.
Brojer, C., Ågren, E. O., Uhlhorn, H., Bernodt, K., Mörner, T., Jansson, D. S., Mattsson, R., Zohari, S., Thorén, P., Berg, M., & Gavier-Widén, D. (2009). Pathology of natural highly pathogenic avian influenza virus H5N1 infection in wild tufted ducks (Aythya fuligula). Journal of Veterinary Diagnostic Investigation, 21, 1663–1670.
Capua, I., & Alexander, D. J. (2009). Avian influenza and Newcastle disease: A field and laboratory manual. Springer-Verlag Italia.
Cattoli, G., Susta, L., Terregino, C., & Brown, C. (2011). Newcastle disease: A review of field recognition and current methods of laboratory detection. Journal of Veterinary Diagnostic Investigation, 23, 637–658.
Chauhan, R. P., & Gordon, M. L. (2022). An overview of influenza A virus genes, protein functions, and replication cycle highlighting important updates. Virus Genes, 58(4), 255–269.
Chauhan, R. P., & Gordon, M. L. (2022). Influenza virus nucleoprotein: Structure, function, and antiviral targeting. Viruses, 14(3), 550.
Creelan, J. L., Graham, D. A., & McCullough, S. J. (2002). Detection and differentiation of pathogenicity of avian paramyxovirus serotype 1 using one-step RT-PCR. Avian Pathology, 31(5), 493–499.
Dewandaru, R. A., Santoso, C., Prayogo, F., & Wibowo, M. H. (2020). Gambaran mikroskopis berbagai organ merpati yang menunjukkan gejala tortikolis dan identifikasi virus penyakit tetelo berdasarkan uji agar gel presipitasi. Jurnal Veteriner, 21(4).
Dewandaru, R. A., et al. (2020). Pathological findings of avian influenza and Newcastle disease in layer chickens. Jurnal Kedokteran Hewan, 14(2), 85–92.
Elton, D., Simpson-Holley, M., Archer, K., Medcalf, L., Hallam, R., McCauley, J., & Digard, P. (1999). Interaction of the influenza virus nucleoprotein with the cellular CRM1-mediated nuclear export pathway. Journal of Virology, 73, 735–745.
Gavin, P. J., & Thomson, R. B. (2003). Review of rapid diagnostic tests for influenza. Clinical and Applied Immunology Reviews, 4, 151–172.
Gayanti, A. N. S., Mahardika, I. G. N. K., Winaya, I. B. O., Suarjana, I. G. K., & Apsari, I. A. P. (2024). Case of avian influenza in broiler chicken at Jatiluwih Village. Buletin Veteriner Udayana, 370–387.
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 protects chickens from lethal H5N1 challenge. Journal of Virology, 81, 150–158.
Ge, S., Zheng, D., Zhao, Y., et al. (2012). Evaluating viral interference between influenza virus and Newcastle disease virus. Journal of Virology, 9, 128.
Haque, M. H., Hossain, M. T., Islam, M. T., Zinnah, M. A., Khan, M. S. R., & Islam, M. A. (2010). Isolation and detection of Newcastle disease virus by RT-PCR. Bangladesh Journal of Veterinary Medicine, 8(2), 87–92.
Hartono, M., Sirat, M. M. P., Santosa, P. E., Siswanto, Ermawati, R., Yudhistira, G. J., Juandita, K. N., Fatmawati, S. T., & Sagala, Y. N. (2021). Vaccination program and the establishment of vaccinator candidates in the context of avian influenza and Newcastle disease prevention in Mandah Village, Lampung Selatan District.
Hoffmann, B., Harder, T., Lange, E., Kalthoff, D., Reimann, I., & Beer, M. (2009). New real-time RT-PCR assays for detection of influenza A viruses. Journal of Virological Methods, 159, 29–36.
Isnawati, R., Putra, I. P. C., Susiani, R. D., Wuryastuti, H., & Wasito, R. (2020). Deteksi virus avian influenza pada ayam pedaging komersial yang disuplementasi water additive. Prosiding SNPBS, 735–743.
Kapczynski, D. R., Afonso, C. L., & Miller, P. J. (2013). Immune responses of poultry to Newcastle disease virus. Developmental and Comparative Immunology, 41, 447–453.
Krzysztof, S., Zenon, M., & Katarzyna, D. (2006). Detection of Newcastle disease virus by RT-PCR. Bulletin of the Veterinary Institute in Pulawy, 50, 3–7.
Lee, M. S., Chang, P. C., Shien, J. H., Cheng, M. C., & Shieh, H. K. (2001). Identification and subtyping of avian influenza viruses by RT-PCR. Journal of Virological Methods, 97, 13–22.
Marson, F. G. S., Suardana, I. B. K., & Nindhia, T. S. (2020). Monitoring titer antibodies post-vaccination of avian influenza subtype H5N1 in duck livestock of Baha Village, Bali.
Mo, J., Segovia, K., Chrzastek, K., Briggs, K., & Kapczynski, D. R. (2024). Morphologic characterization and cytokine response of chicken bone-marrow derived dendritic cells. Frontiers in Immunology, 15, 1374838.
Newcomb, L. L., et al. (2009). Influenza nucleoprotein interactions with polymerase. Journal of Virology, 83, 522–529.
Pantin-Jackwood, M. J., et al. (2023). Pathogenicity in chickens and turkeys of a 2021 US H5N1 virus. Viruses, 15(11), 2273.
Pang, Y., Wang, H., Girshick, T., & Khan, M. I. (2002). Development of multiplex PCR for avian respiratory agents. Avian Diseases, 46, 691–699.
Pattison, M., McMullin, P. F., Bradbury, J. M., & Alexander, D. J. (2008). Poultry diseases (6th ed.). Saunders Elsevier.
Perkins, L. E. L., & Swayne, D. E. (2001). Pathobiology of H5N1 avian influenza virus. Veterinary Pathology, 38, 149–164.
Portela, A., & Digard, P. (2002). The influenza virus nucleoprotein. Journal of General Virology, 83, 723–734.
Pratama, I. G. A. P., et al. (2025). Case study of velogenic Newcastle disease strain. Buletin Veteriner Udayana, 705–720.
Quinn, P. J., et al. (2002). Veterinary microbiology and microbial disease. Blackwell Science.
Rashid, S., et al. (2009). Multiplex PCR for avian respiratory pathogens. Poultry Science, 88, 2526–2531.
Rell, F., Adi, A. A. M., & Mahardika, G. N. K. (2021). Analisis filogeni virus Newcastle disease isolat Bali. Buletin Veteriner Udayana, 13, 67–74.
Ronohardjo, P., et al. (1985). Identification of influenza A virus in Indonesia. Penyakit Hewan, XVII(1), 249–257.
Sanjiwani, K. A. T., et al. (2024). Multiple infections by avian influenza and Newcastle disease virus. Buletin Veteriner Udayana, 1316–1329.
Sari, N. P. D. S., et al. (2024). Newcastle disease and avian influenza in broiler. Veterinary Science and Medicine Journal, 363–375. https://doi.org/10.24843/vsmj.2024.v06.i04.p09
Swayne, D. E., & Pantin-Jackwood, M. (2008). Pathobiology of avian influenza virus infections. Blackwell Publishing.
Wasito, R., et al. (2014). Detection of pathogenic H5 and H7 influenza A viruses. Biochemical and Biotechnology Research, 2(2), 27–31.
Widianingrum, D. C., et al. (2022). Penyakit chronic respiratory disease (CRD). Jurnal Sain Veteriner, 40(2), 221–224.
Wise, M. G., et al. (2004). RNA polymerase gene analysis of Newcastle disease virus. Virus Research, 104(1), 71–80.
Wulandari, I., et al. (2021). Karakterisasi molekuler protein HN virus Newcastle disease. Jurnal Medik Veteriner, 4(2), 231–242.
Wuryastuty, H., & Wasito, R. (2012). Respiratory avian influenza A (H5N1) in ducks. Jurnal Sain Veteriner, 30(1), 1–7.
Wuryastuty, H., & Wasito, R. (2013). Molecular identification of avian influenza virus in house flies. Jurnal Sain Veteriner, 31(1), 1–7.
Zhang, S., et al. (2025). Eco-epidemiological mechanisms of avian influenza and Newcastle virus co-infection. Poultry Science, 105779.
Zou, S. (1997). Genetic screening for influenza virus variants. Journal of Clinical Microbiology, 35, 2623–2627.
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