Korean Journal of Veterinary Research (대한수의학회지)

The Korean Society of Veterinary Science (대한수의학회)
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Production of recombinant nucleocapsid protein of Newcastle disease virus in Escherichia coli for a diagnostic ELISA

  • Kim, Hyun-Il (Lab of Clinical Pathology, Institute of Cheilbio) ;
  • Park, Kyoung-Phil (Lab of Clinical Pathology, Institute of Cheilbio) ;
  • Park, Chan-Hee (Lab of Clinical Pathology, Institute of Cheilbio) ;
  • Cho, Hyun-Ah (Lab of Clinical Pathology, Institute of Cheilbio) ;
  • Yang, Ho-Suk (Lab of Clinical Pathology, Institute of Cheilbio) ;
  • Hahn, Tae-Wook (School of Veterinary Medicine, Kangwon National University)
  • Accepted : 2009.03.26
  • Published : 2009.03.30
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Abstract

Transmission of avian viruses both bird-to-bird and from birds to non-avian species is a major health concern. Newcastle disease virus (NDV) is an economically important avian virus that poses substantial risks to the poultry industry. Rapid and sensitive diagnostic methods, such as the enzymelinked immunosorbent assay (ELISA), are required to track such infections. To develop an ELISA for detecting anti-NDV antibody in avian sera, the nucleocapsid protein (NCP) gene of the NDV La Sota strain was cloned and expressed in Escherichia coli and the 513-amino acid recombinant NCP was purified by Ni-NTA affinity chromatography. To evaluate its ability to replace NDV whole virus antigen as a coating antigen, NCP-coated and whole NDV-coated ELISAs were tested and compared using a panel of NDV positive antisera from chickens. Results using purified NCP were highly correlated with those obtained using whole NDV (r= 0.927), demonstrating that recombinant NCP expressed in Escherichia coli is a suitable substitute antigen for whole NDV in a diagnostic ELISA.

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References

  1. Alexender DJ. Newcastle disease and other avianparamyxoviridae infections. In: Calnek BW, Barnes HJ,Beard CW, McDougald LR, Saif YM (eds.). Diseases of Poultry. 10th ed. pp. 541-569, Iowa State University Press, Ames, 1997
  2. Choppin PW, Compans RW. Reproduction ofparamyxoviruses. In: Fraenkel-Conrat H, Wagner RR(eds.). Comprehensive Virology. Vol. 4. pp. 95-178,Plenum Press, New York, 1975
  3. Errington W, Steward M, Emmerson PT. Adiagnostic immunoassay for Newcastle disease virusbased on the nucleocapsid protein expressed by arecombinant baculovirus. J Virol Methods 1995, 55,357-365 https://doi.org/10.1016/0166-0934(95)00074-7
  4. Laemmli UK. Cleavage of structural proteins duringthe assembly of the head of bacteriophage T4. Nature 1970, 227, 680-685 https://doi.org/10.1038/227680a0
  5. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951, 193, 265-275
  6. Makkay AM, Krell PJ, Nagy E. Antibody detectionbaseddifferential ELISA for NDV-infected orvaccinated chickens versus NDV HN-subunit vaccinatedchickens. Vet Microbiol 1999, 66, 209-222 https://doi.org/10.1016/S0378-1135(99)00016-4
  7. Mebatsion T, Koolen MJ, de Vaan LT, de Haas N,Braber M, Romer-Oberdorfer A, van den Elzen P,van der Marel P. Newcastle disease virus (NDV)marker vaccine: an immunodominant epitope on thenucleoprotein gene of NDV can be deleted or replacedby a foreign epitope. J Virol 2002, 76, 10138-10146 https://doi.org/10.1128/JVI.76.20.10138-10146.2002
  8. Peeters BP, de Leeuw OS, Verstegen I, Koch G,Gielkens AL. Generation of a recombinant chimericNewcastle disease virus vaccine that allows serologicaldifferentiation between vaccinated and infectedanimals. Vaccine 2001, 19, 1616-1627 https://doi.org/10.1016/S0264-410X(00)00419-9
  9. Snyder DB, Marquardt WW, Mallinson ET, RussekE. Rapid serological profiling by enzyme-linkedimmunosorbent assay. I. Measurement of antibodyactivity titer against Newcastle disease virus in a singleserum dilution. Avian Dis 1983, 27, 161-170 https://doi.org/10.2307/1590381
  10. Tan YP, Ling TC, Tan WS, Yusoff K, Tey BT. Recovery of histidine-tagged nucleocapsid protein ofNewcastle disease virus using immobilised metalaffinity chromatography. Process Biochem 2006, 41,874-881 https://doi.org/10.1016/j.procbio.2005.11.003
  11. Tan YP, Ling TC, Yusoff K, Tan WS, Tey BT.Comparative evaluation of three purification methodsfor the nucleocapsid protein of Newcastle disease virusfrom Escherichia coli homogenates. J Microbiol 2005,43, 295-300
  12. Towbin H, Staehelin T, Gordon J. Electrophoretictransfer of proteins from polyacrylamide gels tonitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979, 76, 4350-4354 https://doi.org/10.1073/pnas.76.9.4350
  13. Warnes A, Fooks AR, Stephenson JR. Production ofmeasles nucleoprotein in different expression systemsand its use as a diagnostic reagent. J Virol Methods1994, 49, 257-268 https://doi.org/10.1016/0166-0934(94)90141-4
  14. Williams R, Boshoff CH, Verwoerd D, Schoeman M,van Wyk A, Gerdes TH, Roos K. Detection ofantibodies to Newcastle disease virus in ostriches(Struthio camelus) by an indirect ELISA. Avian Dis1997, 41, 864-869 https://doi.org/10.2307/1592340
  15. Yusoff K, Tan WS. Newcastle disease virus: macromoleculesand opportunities. Avian Pathol 2001, 30,439-455 https://doi.org/10.1080/03079450120078626
  16. Zhou EM, Chan M, Heckert RA, Riva J, CantinMF. Evaluation of a competitive ELISA for detectionof antibodies against avian influenza virus nucleoprotein.Avian Dis 1998, 42, 517-522 https://doi.org/10.2307/1592678