Development of a one-run real-time PCR detection system for pathogens associated with poultry infectious diseases
Infectious diseases are an important issue in the poultry industry, requiring early diagnosis and countermeasures. To address this, we present a system based on TaqMan real-time PCR to detect pathogen genome in specimens collected from chickens. We designed 12 primer–probe sets for pathogens causing...
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| Published in | Journal of Veterinary Medical Science Vol. 85; no. 4; pp. 407 - 411 |
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| Main Authors | , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Japan
JAPANESE SOCIETY OF VETERINARY SCIENCE
2023
Japan Science and Technology Agency The Japanese Society of Veterinary Science |
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| Abstract | Infectious diseases are an important issue in the poultry industry, requiring early diagnosis and countermeasures. To address this, we present a system based on TaqMan real-time PCR to detect pathogen genome in specimens collected from chickens. We designed 12 primer–probe sets for pathogens causing respiratory or systemic symptoms. In field samples, we detected three viruses, including DNA and RNA viruses, and three bacteria. The chicken anemia virus and Avibacterium paragallinarum were detected only in young and laying hens, respectively. Bacteria were detected only in throat swabs, and gallid alphaherpesvirus 2 was detected in different specimens at each developmental stage. Our novel TaqMan real-time PCR system effectively detects pathogen’s gene in chickens, while taking age into account. |
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| AbstractList | Infectious diseases are an important issue in the poultry industry, requiring early diagnosis and countermeasures. To address this, we present a system based on TaqMan real-time PCR to detect pathogen genome in specimens collected from chickens. We designed 12 primer–probe sets for pathogens causing respiratory or systemic symptoms. In field samples, we detected three viruses, including DNA and RNA viruses, and three bacteria. The chicken anemia virus and Avibacterium paragallinarum were detected only in young and laying hens, respectively. Bacteria were detected only in throat swabs, and gallid alphaherpesvirus 2 was detected in different specimens at each developmental stage. Our novel TaqMan real-time PCR system effectively detects pathogen’s gene in chickens, while taking age into account. Infectious diseases are an important issue in the poultry industry, requiring early diagnosis and countermeasures. To address this, we present a system based on TaqMan real-time PCR to detect pathogen genome in specimens collected from chickens. We designed 12 primer-probe sets for pathogens causing respiratory or systemic symptoms. In field samples, we detected three viruses, including DNA and RNA viruses, and three bacteria. The chicken anemia virus and Avibacterium paragallinarum were detected only in young and laying hens, respectively. Bacteria were detected only in throat swabs, and gallid alphaherpesvirus 2 was detected in different specimens at each developmental stage. Our novel TaqMan real-time PCR system effectively detects pathogen's gene in chickens, while taking age into account.Infectious diseases are an important issue in the poultry industry, requiring early diagnosis and countermeasures. To address this, we present a system based on TaqMan real-time PCR to detect pathogen genome in specimens collected from chickens. We designed 12 primer-probe sets for pathogens causing respiratory or systemic symptoms. In field samples, we detected three viruses, including DNA and RNA viruses, and three bacteria. The chicken anemia virus and Avibacterium paragallinarum were detected only in young and laying hens, respectively. Bacteria were detected only in throat swabs, and gallid alphaherpesvirus 2 was detected in different specimens at each developmental stage. Our novel TaqMan real-time PCR system effectively detects pathogen's gene in chickens, while taking age into account. Infectious diseases are an important issue in the poultry industry, requiring early diagnosis and countermeasures. To address this, we present a system based on TaqMan real-time PCR to detect pathogen genome in specimens collected from chickens. We designed 12 primer–probe sets for pathogens causing respiratory or systemic symptoms. In field samples, we detected three viruses, including DNA and RNA viruses, and three bacteria. The chicken anemia virus and Avibacterium paragallinarum were detected only in young and laying hens, respectively. Bacteria were detected only in throat swabs, and gallid alphaherpesvirus 2 was detected in different specimens at each developmental stage. Our novel TaqMan real-time PCR system effectively detects pathogen’s gene in chickens, while taking age into account. |
| ArticleNumber | 22-0482 |
| Author | KAWAHARA, Fumiya NUNOMURA, Yuka MIZUTANI, Tetsuya TAKEMAE, Hitoshi SHIBANUMA, Takuya OBA, Mami |
| Author_xml | – sequence: 1 fullname: SHIBANUMA, Takuya organization: Center for Infectious Disease Epidemiology and Prevention Research, Tokyo University of Agriculture and Technology, Tokyo, Japan – sequence: 2 fullname: NUNOMURA, Yuka organization: Center for Infectious Disease Epidemiology and Prevention Research, Tokyo University of Agriculture and Technology, Tokyo, Japan – sequence: 3 fullname: OBA, Mami organization: Center for Infectious Disease Epidemiology and Prevention Research, Tokyo University of Agriculture and Technology, Tokyo, Japan – sequence: 4 fullname: KAWAHARA, Fumiya organization: Center for Infectious Disease Epidemiology and Prevention Research, Tokyo University of Agriculture and Technology, Tokyo, Japan – sequence: 5 fullname: MIZUTANI, Tetsuya organization: Center for Infectious Disease Epidemiology and Prevention Research, Tokyo University of Agriculture and Technology, Tokyo, Japan – sequence: 6 fullname: TAKEMAE, Hitoshi organization: Center for Infectious Disease Epidemiology and Prevention Research, Tokyo University of Agriculture and Technology, Tokyo, Japan |
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| References | 17. Rahpaya SS, Tsuchiaka S, Kishimoto M, Oba M, Katayama Y, Nunomura Y, Kokawa S, Kimura T, Kobayashi A, Kirino Y, Okabayashi T, Nonaka N, Mekata H, Aoki H, Shiokawa M, Umetsu M, Morita T, Hasebe A, Otsu K, Asai T, Yamaguchi T, Makino S, Murata Y, Abi AJ, Omatsu T, Mizutani T. 2018. Dembo polymerase chain reaction technique for detection of bovine abortion, diarrhea, and respiratory disease complex infectious agents in potential vectors and reservoirs. J Vet Sci 19: 350–357. 25. Techera C, Tomás G, Panzera Y, Banda A, Perbolianachis P, Pérez R, Marandino A. 2019. Development of real-time PCR assays for single and simultaneous detection of infectious bursal disease virus and chicken anemia virus. Mol Cell Probes 43: 58–63. 7. Ikezawa M, Goryo M, Sasaki J, Haridy M, Okada K. 2010. Late Marek’s disease in adult chickens inoculated with virulent Marek’s disease virus. J Vet Med Sci 72: 1539–1545. 30. Witter RL, Sharma JM, Solomon JJ, Champion LR. 1973. An age-related resistance of chickens to Marek’s disease: some preliminary observations. Avian Pathol 2: 43–54. 20. Sharma JM, Witter RL, Burmester BR. 1973. Pathogenesis of Marek’s disease in old chickens: lesion regression as the basis for age-related resistance. Infect Immun 8: 715–724. 8. Jayasundara JMKGK, Walkden-Brown SW, Katz ME, Islam AFMF, Renz KG, McNally J, Hunt PW. 2017. Pathogenicity, tissue distribution, shedding and environmental detection of two strains of IBDV following infection of chickens at 0 and 14 days of age. Avian Pathol 46: 242–255. 2. Clothier KA, Torain A, Reinl S. 2019. Surveillance for Avibacterium paragallinarum in autopsy cases of birds from small chicken flocks using a real-time PCR assay. J Vet Diagn Invest 31: 364–367. 12. Laamiri N, Aouini R, Marnissi B, Ghram A, Hmila I. 2018. A multiplex real-time RT-PCR for simultaneous detection of four most common avian respiratory viruses. Virology 515: 29–37. 14. Mase M, Tsukamoto K, Imai K, Yamaguchi S. 2004. Phylogenetic analysis of avian infectious bronchitis virus strains isolated in Japan. Arch Virol 149: 2069–2078. 21. Sprygin AV, Andreychuk DB, Kolotilov AN, Volkov MS, Runina IA, Mudrak NS, Borisov AV, Irza VN, Drygin VV, Perevozchikova NA. 2010. Development of a duplex real-time TaqMan PCR assay with an internal control for the detection of Mycoplasma gallisepticum and Mycoplasma synoviae in clinical samples from commercial and backyard poultry. Avian Pathol 39: 99–109. 28. Wernicki A, Nowaczek A, Urban-Chmiel R. 2017. Bacteriophage therapy to combat bacterial infections in poultry. Virol J 14: 179. 11. Kiss I, Matiz K, Kaszanyitzky E, Chávez Y, Johansson KE. 1997. Detection and identification of avian mycoplasmas by polymerase chain reaction and restriction fragment length polymorphism assay. Vet Microbiol 58: 23–30. 13. Markowski-Grimsrud CJ, Schat KA. 2003. Infection with chicken anaemia virus impairs the generation of pathogen-specific cytotoxic T lymphocytes. Immunology 109: 283–294. 29. Wernike K, Hoffmann B, Kalthoff D, König P, Beer M. 2011. Development and validation of a triplex real-time PCR assay for the rapid detection and differentiation of wild-type and glycoprotein E-deleted vaccine strains of Bovine herpesvirus type 1. J Virol Methods 174: 77–84. 15. Miller MM, Jarosinski KW, Schat KA. 2005. Positive and negative regulation of chicken anemia virus transcription. J Virol 79: 2859–2868. 19. Schat KA, Van Santen V. 2008. Chicken infectious anemia. pp. 211–235. In: Diseases of Poultry (Saif YM, Fadly AM, Glisson JR, McDougald LR, Nolan LK, Swayne DE eds.), Blackwell Publishing, Ames. 16. Norup LR, Jensen KH, Jørgensen E, Sørensen P, Juul-Madsen HR. 2008. Effect of mild heat stress and mild infection pressure on immune responses to an E. coli infection in chickens. Animal 2: 265–274. 31. Yuasa N, Noguchi T, Furuta K, Yoshida I. 1980. Maternal antibody and its effect on the susceptibility of chicks to chicken anemia agent. Avian Dis 24: 197–201. 9. Kennedy DA, Cairns C, Jones MJ, Bell AS, Salathé RM, Baigent SJ, Nair VK, Dunn PA, Read AF. 2017. Industry-wide surveillance of Marek’s disease virus on commercial poultry farms. Avian Dis 61: 153–164. 6. Ignjatović J, Sapats S. 2000. Avian infectious bronchitis virus. Rev Sci Tech 19: 493–508. 1. Baigent SJ, Nair VK, Le Galludec H. 2016. Real-time PCR for differential quantification of CVI988 vaccine virus and virulent strains of Marek’s disease virus. J Virol Methods 233: 23–36. 18. Rozins C, Day T, Greenhalgh S. 2019. Managing Marek’s disease in the egg industry. Epidemics 27: 52–58. 10. Kishimoto M, Tsuchiaka S, Rahpaya SS, Hasebe A, Otsu K, Sugimura S, Kobayashi S, Komatsu N, Nagai M, Omatsu T, Naoi Y, Sano K, Okazaki-Terashima S, Oba M, Katayama Y, Sato R, Asai T, Mizutani T. 2017. Development of a one-run real-time PCR detection system for pathogens associated with bovine respiratory disease complex. J Vet Med Sci 79: 517–523. 22. Stipkovits L, Kempf I. 1996. Mycoplasmoses in poultry. Rev Sci Tech 15: 1495–1525. 23. Sun F, Ferro PJ, Lupiani B, Kahl J, Morrow ME, Flanagan JP, Estevez C, Clavijo A. 2011. A duplex real-time polymerase chain reaction assay for the simultaneous detection of long terminal repeat regions and envelope protein gene sequences of Reticuloendotheliosis virus in avian blood samples. J Vet Diagn Invest 23: 937–941. 24. Sunaga F, Tsuchiaka S, Kishimoto M, Aoki H, Kakinoki M, Kure K, Okumura H, Okumura M, Okumura A, Nagai M, Omatsu T, Mizutani T. 2020. Development of a one-run real-time PCR detection system for pathogens associated with porcine respiratory diseases. J Vet Med Sci 82: 217–223. 4. Glisson JR. 1998. Bacterial respiratory disease of poultry. Poult Sci 77: 1139–1142. 27. Untergasser A, Nijveen H, Rao X, Bisseling T, Geurts R, Leunissen JA. 2007. Primer3Plus, an enhanced web interface to Primer3. Nucleic Acids Res 35: W71-4. 26. Tsuchiaka S, Masuda T, Sugimura S, Kobayashi S, Komatsu N, Nagai M, Omatsu T, Furuya T, Oba M, Katayama Y, Kanda S, Yokoyama T, Mizutani T. 2016. Development of a novel detection system for microbes from bovine diarrhea by real-time PCR. J Vet Med Sci 78: 383–389. 3. Davidson I, Raibstein I, Altory A. 2015. Differential diagnosis of fowlpox and infectious laryngotracheitis viruses in chicken diphtheritic manifestations by mono and duplex real-time polymerase chain reaction. Avian Pathol 44: 1–4. 5. Guo M, Liu D, Chen X, Wu Y, Zhang X. 2022. Pathogenicity and innate response to Avibacterium paragallinarum in chickens. Poult Sci 101: 101523. 22 23 24 25 26 27 28 29 30 31 10 11 12 13 14 15 16 17 18 19 1 2 3 4 5 6 7 8 9 20 21 |
| References_xml | – reference: 20. Sharma JM, Witter RL, Burmester BR. 1973. Pathogenesis of Marek’s disease in old chickens: lesion regression as the basis for age-related resistance. Infect Immun 8: 715–724. – reference: 2. Clothier KA, Torain A, Reinl S. 2019. Surveillance for Avibacterium paragallinarum in autopsy cases of birds from small chicken flocks using a real-time PCR assay. J Vet Diagn Invest 31: 364–367. – reference: 5. Guo M, Liu D, Chen X, Wu Y, Zhang X. 2022. Pathogenicity and innate response to Avibacterium paragallinarum in chickens. Poult Sci 101: 101523. – reference: 19. Schat KA, Van Santen V. 2008. Chicken infectious anemia. pp. 211–235. In: Diseases of Poultry (Saif YM, Fadly AM, Glisson JR, McDougald LR, Nolan LK, Swayne DE eds.), Blackwell Publishing, Ames. – reference: 27. Untergasser A, Nijveen H, Rao X, Bisseling T, Geurts R, Leunissen JA. 2007. Primer3Plus, an enhanced web interface to Primer3. Nucleic Acids Res 35: W71-4. – reference: 24. Sunaga F, Tsuchiaka S, Kishimoto M, Aoki H, Kakinoki M, Kure K, Okumura H, Okumura M, Okumura A, Nagai M, Omatsu T, Mizutani T. 2020. Development of a one-run real-time PCR detection system for pathogens associated with porcine respiratory diseases. J Vet Med Sci 82: 217–223. – reference: 18. Rozins C, Day T, Greenhalgh S. 2019. Managing Marek’s disease in the egg industry. Epidemics 27: 52–58. – reference: 25. Techera C, Tomás G, Panzera Y, Banda A, Perbolianachis P, Pérez R, Marandino A. 2019. Development of real-time PCR assays for single and simultaneous detection of infectious bursal disease virus and chicken anemia virus. Mol Cell Probes 43: 58–63. – reference: 15. Miller MM, Jarosinski KW, Schat KA. 2005. Positive and negative regulation of chicken anemia virus transcription. J Virol 79: 2859–2868. – reference: 11. Kiss I, Matiz K, Kaszanyitzky E, Chávez Y, Johansson KE. 1997. Detection and identification of avian mycoplasmas by polymerase chain reaction and restriction fragment length polymorphism assay. Vet Microbiol 58: 23–30. – reference: 17. Rahpaya SS, Tsuchiaka S, Kishimoto M, Oba M, Katayama Y, Nunomura Y, Kokawa S, Kimura T, Kobayashi A, Kirino Y, Okabayashi T, Nonaka N, Mekata H, Aoki H, Shiokawa M, Umetsu M, Morita T, Hasebe A, Otsu K, Asai T, Yamaguchi T, Makino S, Murata Y, Abi AJ, Omatsu T, Mizutani T. 2018. Dembo polymerase chain reaction technique for detection of bovine abortion, diarrhea, and respiratory disease complex infectious agents in potential vectors and reservoirs. J Vet Sci 19: 350–357. – reference: 28. Wernicki A, Nowaczek A, Urban-Chmiel R. 2017. Bacteriophage therapy to combat bacterial infections in poultry. Virol J 14: 179. – reference: 13. Markowski-Grimsrud CJ, Schat KA. 2003. Infection with chicken anaemia virus impairs the generation of pathogen-specific cytotoxic T lymphocytes. Immunology 109: 283–294. – reference: 12. Laamiri N, Aouini R, Marnissi B, Ghram A, Hmila I. 2018. A multiplex real-time RT-PCR for simultaneous detection of four most common avian respiratory viruses. Virology 515: 29–37. – reference: 9. Kennedy DA, Cairns C, Jones MJ, Bell AS, Salathé RM, Baigent SJ, Nair VK, Dunn PA, Read AF. 2017. Industry-wide surveillance of Marek’s disease virus on commercial poultry farms. Avian Dis 61: 153–164. – reference: 16. Norup LR, Jensen KH, Jørgensen E, Sørensen P, Juul-Madsen HR. 2008. Effect of mild heat stress and mild infection pressure on immune responses to an E. coli infection in chickens. Animal 2: 265–274. – reference: 22. Stipkovits L, Kempf I. 1996. Mycoplasmoses in poultry. Rev Sci Tech 15: 1495–1525. – reference: 8. Jayasundara JMKGK, Walkden-Brown SW, Katz ME, Islam AFMF, Renz KG, McNally J, Hunt PW. 2017. Pathogenicity, tissue distribution, shedding and environmental detection of two strains of IBDV following infection of chickens at 0 and 14 days of age. Avian Pathol 46: 242–255. – reference: 14. Mase M, Tsukamoto K, Imai K, Yamaguchi S. 2004. Phylogenetic analysis of avian infectious bronchitis virus strains isolated in Japan. Arch Virol 149: 2069–2078. – reference: 4. Glisson JR. 1998. Bacterial respiratory disease of poultry. Poult Sci 77: 1139–1142. – reference: 21. Sprygin AV, Andreychuk DB, Kolotilov AN, Volkov MS, Runina IA, Mudrak NS, Borisov AV, Irza VN, Drygin VV, Perevozchikova NA. 2010. Development of a duplex real-time TaqMan PCR assay with an internal control for the detection of Mycoplasma gallisepticum and Mycoplasma synoviae in clinical samples from commercial and backyard poultry. Avian Pathol 39: 99–109. – reference: 29. Wernike K, Hoffmann B, Kalthoff D, König P, Beer M. 2011. Development and validation of a triplex real-time PCR assay for the rapid detection and differentiation of wild-type and glycoprotein E-deleted vaccine strains of Bovine herpesvirus type 1. J Virol Methods 174: 77–84. – reference: 31. Yuasa N, Noguchi T, Furuta K, Yoshida I. 1980. Maternal antibody and its effect on the susceptibility of chicks to chicken anemia agent. Avian Dis 24: 197–201. – reference: 6. Ignjatović J, Sapats S. 2000. Avian infectious bronchitis virus. Rev Sci Tech 19: 493–508. – reference: 10. Kishimoto M, Tsuchiaka S, Rahpaya SS, Hasebe A, Otsu K, Sugimura S, Kobayashi S, Komatsu N, Nagai M, Omatsu T, Naoi Y, Sano K, Okazaki-Terashima S, Oba M, Katayama Y, Sato R, Asai T, Mizutani T. 2017. Development of a one-run real-time PCR detection system for pathogens associated with bovine respiratory disease complex. J Vet Med Sci 79: 517–523. – reference: 23. Sun F, Ferro PJ, Lupiani B, Kahl J, Morrow ME, Flanagan JP, Estevez C, Clavijo A. 2011. A duplex real-time polymerase chain reaction assay for the simultaneous detection of long terminal repeat regions and envelope protein gene sequences of Reticuloendotheliosis virus in avian blood samples. J Vet Diagn Invest 23: 937–941. – reference: 7. Ikezawa M, Goryo M, Sasaki J, Haridy M, Okada K. 2010. Late Marek’s disease in adult chickens inoculated with virulent Marek’s disease virus. J Vet Med Sci 72: 1539–1545. – reference: 26. Tsuchiaka S, Masuda T, Sugimura S, Kobayashi S, Komatsu N, Nagai M, Omatsu T, Furuya T, Oba M, Katayama Y, Kanda S, Yokoyama T, Mizutani T. 2016. Development of a novel detection system for microbes from bovine diarrhea by real-time PCR. J Vet Med Sci 78: 383–389. – reference: 30. Witter RL, Sharma JM, Solomon JJ, Champion LR. 1973. An age-related resistance of chickens to Marek’s disease: some preliminary observations. Avian Pathol 2: 43–54. – reference: 1. Baigent SJ, Nair VK, Le Galludec H. 2016. Real-time PCR for differential quantification of CVI988 vaccine virus and virulent strains of Marek’s disease virus. J Virol Methods 233: 23–36. – reference: 3. Davidson I, Raibstein I, Altory A. 2015. Differential diagnosis of fowlpox and infectious laryngotracheitis viruses in chicken diphtheritic manifestations by mono and duplex real-time polymerase chain reaction. 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| Snippet | Infectious diseases are an important issue in the poultry industry, requiring early diagnosis and countermeasures. To address this, we present a system based... Infectious diseases are an important issue in the poultry industry, requiring early diagnosis and countermeasures. To address this, we present a system based... |
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| SubjectTerms | Animals Avian Pathology chicken Chickens - microbiology Communicable Diseases - veterinary DNA viruses Female Genomes infectious disease Infectious diseases pathogen detection system Pathogens Polymerase chain reaction Poultry Poultry Diseases real-time PCR Real-Time Polymerase Chain Reaction - veterinary RNA Viruses |
| Title | Development of a one-run real-time PCR detection system for pathogens associated with poultry infectious diseases |
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