J Vet Sci.  2013 Sep;14(3):291-297. 10.4142/jvs.2013.14.3.291.

Preparation and diagnostic utility of a hemagglutination inhibition test antigen derived from the baculovirus-expressed hemagglutinin-neuraminidase protein gene of Newcastle disease virus

• Choi KS
• Kye SJ
• Jeon WJ
• Park MJ
• Kim S
• Seul HJ
• Kwon JH

• KMID: 1705559
• DOI: http://doi.org/10.4142/jvs.2013.14.3.291

Abstract

A recombinant hemagglutinin-neuraminidase (rHN) protein from Newcastle disease virus (NDV) with hemagglutination (HA) activity was expressed in Spodoptera frugiperda cells using a baculovirus expression system. The rHN protein extracted from infected cells was used as an antigen in a hemagglutination inhibition (HI) test for the detection and titration of NDV-specific antibodies present in chicken sera. The rHN antigen produced high HA titers of 2(13) per 25 microL, which were similar to those of the NDV antigen produced using chicken eggs, and it remained stable without significant loss of the HA activity for at least 12 weeks at 4degrees C. The rHN-based HI assay specifically detected NDV antibodies, but not the sera of other avian pathogens, with a specificity and sensitivity of 100% and 98.0%, respectively, in known positive and negative chicken sera (n = 430). Compared with an NDV-based HI assay, the rHN-based HI assay had a relative sensitivity and specificity of 96.1% and 95.5%, respectively, when applied to field chicken sera. The HI titers of the rHN-based HI assay were highly correlated with those in an NDV-based HI assay (r = 0.927). Overall, these results indicate that rHN protein provides a useful alternative to NDV antigen in HI assays.

Keyword

antibody detection; hemagglutinin-neuraminidase protein; hemagglutination inhibition; Newcastle disease virus

MeSH Terms

Animals
Antibodies, Viral/*blood
Antigens, Viral/*diagnostic use/genetics/metabolism
Baculoviridae/genetics
Chickens
HN Protein/*diagnostic use/genetics/metabolism
Hemagglutination Inhibition Tests/*methods/veterinary
Newcastle Disease/*diagnosis/immunology/virology
Newcastle disease virus/genetics/*immunology/metabolism
Poultry Diseases/*diagnosis/immunology/virology
Recombinant Proteins/diagnostic use/genetics/metabolism
Sf9 Cells
Spodoptera
Antibodies, Viral
Antigens, Viral
HN Protein
Recombinant Proteins

Figure

• 

  Fig. 1 Construction of an expression vector containing the entire open reading frame of the hemagglutinin-neuraminidase (HN) gene from Newcastle disease virus (NDV).

• 

  Fig. 2 Western blot analysis (A) and hemagglutination (HA) titer (B) of cell lysates from recombinant baculovirus-infected cells. The arrow in Fig. 1A indicates recombinant hemagglutinin-neuraminidase (rHN) proteins binding to NDV antibodies. The HA titer was determined as the reciprocal of the highest dilution with HA activity. M: protein molecular weight marker; Lane 1: infected cell lysate; Lane 2; normal cell lysate.

• 

  Fig. 3 Correlation between serum hemagglutination inhibition (HI) titers obtained using the rHN-based HI and NDV-based HI assays. Test sera were from healthy commercial chickens (n = 791) in broiler farms where vaccination was performed routinely.

Reference

1. Afonso CL, Miller PJ, Grund C, Koch G, Peeters B, Selleck PW, Srinivas GB. Newcastle disease. OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Aanimals, 2012. 7th ed. Paris: OIE;2012. p. 555–573.

2. Alexander DJ. Newcastle disease and other avian paramyxoviruses. Rev Sci Tech. 2000; 19:443–462.

3. Alexander DJ. Newcastle disease in the European Union 2000 to 2009. Avian Pathol. 2011; 40:547–558.

4. Anderson SC. A note on two laboratory infections with the virus of Newcastle disease. Med J Aust. 1946; 1:371.

5. Beard CW, Hopkins SR, Hammond J. Preparation of Newcastle disease virus hemagglutination-inhibition test antigen. Avian Dis. 1975; 19:692–699.

6. Bose S, Welch BD, Kors CA, Yuan P, Jardetzky TS, Lamb RA. Structure and mutagenesis of the parainfluenza virus 5 hemagglutinin-neuraminidase stalk domain reveals a four-helix bundle and the role of the stalk in fusion promotion. J Virol. 2011; 85:12855–12866.

7. Burnet FM. Human infection with the virus of Newcastle disease of fowl. Med J Aust. 1943; 2:313–314.

8. Cho SH, Kwon HJ, Kim TE, Kim JH, Yoo HS, Kim SJ. Variation of a Newcastle disease virus hemagglutinin-neuraminidase linear epitope. J Clin Microbiol. 2008; 46:1541–1544.

9. Cho SH, Kwon HJ, Kim TE, Kim JH, Yoo HS, Park MH, Park YH, Kim SJ. Characterization of a recombinant Newcastle disease virus vaccine strain. Clin Vaccine Immunol. 2008; 15:1572–1579.

10. Cosset FL, Bouquet JF, Drynda A, Chebloune Y, Rey-Senelonge A, Kohen G, Nigon VM, Desmettre P, Verdier G. Newcastle disease virus (NDV) vaccine based on immunization with avian cells expressing the NDV hemagglutinin-neuraminidase glycoprotein. Virology. 1991; 185:862–866.

11. Dardiri AH, Yates VM, Flanagan TD. The reaction to infection with the B1 strain of Newcastle disease virus in man. Am J Vet Res. 1962; 23:918–921.

12. Dortmans JCFM, Peeters BPH, Koch G. Newcastle disease virus outbreaks: Vaccine mismatch or inadequate application? Vet Microbiol. 2012; 160:17–22.

13. Freymann MW, Bang FB. Human conjunctivitis due to Newcastle virus in the U.S.A. Bull Johns Hopkins Hosp. 1949; 84:409–413.

14. Goebel SJ, Taylor J, Barr BC, Kiehn TE, Castroalaspina HR, Hedvat CV, Rush-Wilson KA, Kelly CD, Davis SW, Samsonoff WA, Hurst KR, Behr MJ, Masters PS. Isolation of avian paramyxovirus 1 from a patient with a lethal case of pneumonia. J Virol. 2007; 81:12709–12714.

15. Gustafson DP, Moses HE. Isolation of Newcastle disease virus from the eye of a human being. J Am Vet Assoc. 1951; 117:1–2.

16. Heckert RA, Nagy E. Evaluation of the hemagglutiniation-inhibition assay using a baculovirus-expressed hemagglutinin-neuraminidase protein for detection of Newcastle disease virus antibodies. J Vet Diagn Invest. 1999; 11:99–102.

17. Hu S, Ma H, Wu Y, Liu W, Wang X, Liu Y, Liu X. A vaccine candidate of attenuated genotype VII Newcastle disease virus generated by reverse genetics. Vaccine. 2009; 27:904–910.

18. Hu S, Wang T, Liu Y, Meng C, Wang X, Wu Y, Liu X. Identification of a variable epitope on the Newcastle disease virus hemagglutinin-neuraminidase protein. Vet Microbiol. 2010; 140:92–97.

19. Huang Z, Panda A, Elankumaran S, Govindarajan D, Rockemann D, Samal SK. The hemagglutinin-neuraminidase protein of Newcastle disease virus determines tropism and virulence. J Virol. 2004; 78:4176–4184.

20. Kapczynski DR, King DJ. Protection of chickens against overt clinical disease and determination of viral shedding following vaccination with commercially available Newcastle disease virus vaccines upon challenge with highly virulent virus from the California 2002 exotic Newcastle disease outbreak. Vaccine. 2005; 23:3424–3433.

21. Lee YJ, Sung HW, Choi JG, Kim JH, Song CS. Molecular epidemiology of Newcastle disease viruses isolated in South Korea using sequencing of the fusion protein cleavage site region and phylogenetic relationships. Avian Pathol. 2004; 33:482–491.

22. Mayo MA. A summary of taxonomic changes recently approved by ICTV. Arch Virol. 2002; 147:1655–1663.

23. Miller PJ, Estevez C, Yu Q, Suarez DL, King DJ. Comparison of viral shedding following vaccination with inactivated and live Newcastle disease vaccines formulated with wild-type and recombinant viruses. Avian Dis. 2009; 53:39–49.

24. Mohan CM, Dey S, Rai A, Kataria JM. Recombinant haemagglutinin neuraminidase antigen-based single serum dilution ELISA for rapid serological profiling of Newcastle disease virus. J Virol Methods. 2006; 138:117–122.

25. Moro de Sousa RL, Montassier HJ, Pinto AA. Detection and quantification of antibodies to Newcastle disease virus in ostrich and rhea sera using a liquid phase blocking enzyme-linked immunosorbent assay. Clin Diagn Lab Immunol. 2000; 7:940–944.

26. Mustaffa-Babjee A, Ibrahima AL, Khim TS. A case of human infection with Newcastle disease virus. Southeast Asian J Trop Med Public Health. 1976; 7:622–624.

27. Nagy E, Derbyshire JB, Dobos P, Krell PJ. Cloning and expression of NDV hemagglutinin-neuraminidase cDNA in a baculovirus expression vector system. Virology. 1990; 176:426–438.

28. Nagy E, Krell PJ, Dulac GC, Derbyshire JB. Vaccination against Newcastle disease with a recombinant baculovirus hemagglutinin-neuraminidase subunit vaccine. Avian Dis. 1991; 35:585–590.

29. Niikura M, Matsuura Y, Hattori M, Onuma M, Mikami T. Characterization of haemagglutinin-neuraminidase glycoprotein of Newcastle disease virus expressed by a recombinant baculovirus. Virus Res. 1991; 20:31–43.

30. Ong HKA, Ali AM, Omar AR, Tan WS, Yusoff K. N-linked glycosylated HN protein of NDV strain AF2240 expressed in baculovirus-infected Sf9 cells. J Biochem Mol Biol Biophys. 1999; 3:147–151.

31. Ong HKA, Ali AM, Omar AR, Yusoff K. Cloning and expression of the HN gene from the velogenic viscerotropic Newcastle disease virus strain AF2240 in Sf9 insect cells. Cytotechnology. 2000; 32:243–251.

32. Shimkin NI. Conjunctival haemorrhage due to an infection of Newcastle virus of fowls in man: laboratory and contact infection. Br J Ophthalmol. 1946; 30:260–264.

33. Swayne DE, King DJ. Avian influenza and Newcastle disease. J Am Vet Med Assoc. 2003; 222:1534–1540.

34. Tan WS, Lau CH, Ng BK, Ibrahim AL, Yusoff K. Nucleotide sequence of the haemagglutinin-neuraminidase (HN) gene of Malaysian heat resistant viscerotropic-velogenic Newcastle disease virus (NDV) strain AF2240. DNA Seq. 1995; 6:47–50.

35. Yan Y, Rout SN, Kim SH, Samal SK. Role of untranslated regions of hemagglutinin-neuraminidase gene in replication and pathogenicity of Newcastle disease virus. J Virol. 2009; 83:5943–5946.

36. Yatom J. An outbreak of conjunctivitis in man associated with the virus of Newcastle disease. Harefuah. 1946; 3:69–70.

37. Yuan P, Swanson KA, Leser GP, Paterson RG, Lamb RA, Jardetzky TS. Structure of the Newcastle disease virus hemagglutinin-neuraminidase (HN) ectodomain reveals a four-helix bundle stalk. Proc Natl Acad Sci U S A. 2011; 108:14920–14925.

38. Zoth SC, Gómez E, Carballeda JM, Carrillo E, Berinstein A. Expression of a secreted version of the hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent. J Vet Diagn Invest. 2011; 23:519–523.