Detection and differentiation of Newcastle disease virus and influenza virus by using duplex real-time PCR

• Authors: Dawid Nidzworski , Edyta Wasilewska , Krzysztof Smietanka , Bogusław Szewczyk , Zenon Minta
• Languages of publication: EN

Abstracts

EN

Newcastle disease virus (NDV), member of the Paramyxoviridae family and avian influenza virus (AIV), member of the Orthomyxoviridae family, are two main avian pathogens causing serious economic problems in poultry health. Both are enveloped, single-stranded, negative-sense RNA viruses and cause similar symptoms, ranging from sub-clinical infections to severe diseases, including decrease in egg production, acute respiratory syndrome, and high mortality. Similar symptoms hinder the differentiation of infection with the two viruses by standard veterinary procedures like clinical examination or necropsy. To overcome this problem, we have developed a new duplex real-time PCR assay for the detection and differentiation of these two viruses. Eighteen NDV strains, fourteen AIV strains, and twelve other (negative control) strains viruses were isolated from allantoic fluids of specific pathogen-free (SPF), embryonated eggs. Four-weeks-old SPF chickens were co-infected with both viruses (NDV - LaSota and AIV - H7N1). Swabs from cloaca and trachea were collected and examined. The results obtained in this study show that by using duplex real-time PCR, it was possible to detect and distinguish both viruses within less than three hours and with high sensitivity, even in case a bird was co-infected. Additionally, the results show the applicability of the real-time PCR assay in laboratory practice for the identification and differentiation of Newcastle disease and influenza A viruses in birds.

Keywords

EN

Newcastle disease virus; influenza virus; detection; differentiation; real-time PCR; duplex

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2013
• Volume: 60
• Issue: 3
• Pages: 475-480

Dates

published

2013

received

2013-05-14

revised

2013-07-24

accepted

2013-09-01

(unknown)

2013-09-13

Contributors

author

Dawid Nidzworski

• Department of Molecular Virology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Gdańsk, Poland

author

Edyta Wasilewska

• Department of Molecular Virology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Gdańsk, Poland

author

Krzysztof Smietanka

• The National Veterinary Research Institute, Puławy, Poland

author

Bogusław Szewczyk

• Department of Molecular Virology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Gdańsk, Poland

author

Zenon Minta

• The National Veterinary Research Institute, Puławy, Poland

References

• Alexander DJ (1993) Orthomyxovirus infections. In Viral Infections of Birds. McFerran JB, McNulty MS, eds, pp 287-316. Elsevier Science, London.
• Beard CW, Hanson RP (1984) Newcastle disease. In Disease of Poultry, 8th edn. Hofstad MS, Barnes HJ, Calnek BW, Reid WM, Yoder HW, eds, pp 452-470. Iowa State University Press, Ames.
• Capua I, Alexander DJ (2002) Avian influenza and human health. Acta Trop 83: 1-6.
• Chen H, Zhang J, Sun D, Zhang D, Cai X, Liu X, Ding Y, Ma L, Yang S, Jin L, Liu Y (2008) Rapid discrimination of H5 and H9 subtypes of avian influenza viruses and Newcastle disease virus by multiplex RT-PCR. Vet Res Commun 32: 491-498.
• Collins MS, Strong I, Alexander DJ (1996) Pathogenicity and phylogenetic evaluation of the variant Newcastle disease viruses termed 'pigeon PMV-1 viruses' based on the nucleotide sequence of the fusion protein gene. Arch Virol 141: 635-647.
• Ke GM, Cheng HL, Ke LY, Ji WT, Chulu JL, Liao MH, Chang TJ, Liu HJ (2006) Development of a quantitative Light Cycler real-time RT-PCR for detection of avian reovirus. J Virol Methods 133: 6-13.
• Lamb RA, Collins PL, Kolakofsky D, Melero JA, Nagai Y, Oldstone MBA, Pringle CR, Rima BK (2005) Family paramyxoviridae,. In Virus Taxonomy, VIIIth Report of the International Committee on Taxonomy of Viruses. Fauquet CM, Mayo MA, Maniloff J, Desselberger U, Ball LA, eds, pp 655-668. Elsevier Academic Press, San Diego.
• Mia Kim L, Suarez DL, Afonso CL (2008) Detection of a broad range of class I and II Newcastle disease viruses using a multiplex real time reverse transcription polymerase chain reaction assay. J Vet Diagn Invest 20: 414-425.
• Nagai Y, Klenk HD, Rott R (1976) Proteolytic cleavage of the viral glycoproteins and its significance for the virulence of Newcastle disease virus. Virology 72: 494-508.
• Nidzworski D, Rabalski Ł, Gromadzka B (2011) Detection and differentiation of virulent and avirulent strains of Newcastle Disease Virus by real-time PCR. J Virol Methods 173: 144-149.
• OIE (2009) Manual of diagnostic tests and vaccines for terrestrial animals: mammals, birds and bees. In: Biological Standards Commission, World Organization for Animal Health, pp. 576-589. Paris.
• Panda A, Huang Z, Elankumaran S, Rockemann DD, Samal SK (2004) Role of fusion protein cleavage site in the virulence of Newcastle disease virus. Microb Pathog 36: 1-10.
• Rashid S, Naeem K, Ahmed Z, Saddique N, Abbas MA, Malik SA (2009) Multiplex polymerase chain reaction for the detection and differentiation of avian influenza viruses and other poultry respiratory pathogens. Poult Sci 88: 2526-2531.
• Rott R, Klenk HD (1988) Molecular basis of infectivity and pathogenicity of Newcastle disease virus. In Newcastle Disease. Alexander DJ, eds, pp 98-112. Kluwer Academic Publishers, Boston, MA.
• Seal BS, King DJ, Bennet JD (1995) Characterization of Newcastle disease virus isolates by reverse transcription PCR coupled to direct nucleotide sequencing and development of sequence database for pathotype prediction and molecular epidemiological analysis. J Clin Microbiol 33: 2624-2630.
• Seal BS, King DJ, Sellers HS (2000) The avian response to Newcastle disease virus. Dev Comp Immunol 24: 257-268.
• Spackman E, Senne DA, Myers TJ, Bulaga LL, Garber LP, Perdue ML, Lohman K, Daum LT, Suarez DL (2002) Development of a Real-Time Reverse Transcriptase PCR Assay for Type A Influenza Virus and the Avian H5 and H7 Hemagglutinin Subtypes. J Clin Microbiol 40: 3256-3260.
• Suarez DL, Schultz-Cherry S (2000) Immunology of avian influenza virus: a review. Dev Comp Immunol 24: 269-283.
• Tang Q, Wang J, Bao J, Sun H, Sun Y, Liu J, Pu J (2012) A multiplex RT-PCR assay for detection and differentiation of avian H3, H5, and H9 subtype influenza viruses and Newcastle disease viruses. J Virol Methods 181: 164-169.