Browse > Article
http://dx.doi.org/10.14405/kjvr.2021.61.e19

Expression of the VP2 protein of feline panleukopenia virus in insect cells and use thereof in a hemagglutination inhibition assay  

Yang, Dong-Kun (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs)
Park, Yeseul (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs)
Park, Yu-Ri (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs)
Yoo, Jae Young (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs)
An, Sungjun (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs)
Park, Jungwon (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs)
Hyun, Bang-Hun (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs)
Publication Information
Korean Journal of Veterinary Research / v.61, no.2, 2021 , pp. 19.1-19.7 More about this Journal
Abstract
Feline panleukopenia virus (FPV) causes leukopenia and severe hemorrhagic diarrhea, killing 50% of naturally infected cats. Although intact FPV can serve as an antigen in the hemagglutination inhibition (HI) test, an accidental laboratory-mediated infection is concern. A non-infectious diagnostic reagent is required for the HI test. Here, we expressed the viral protein 2 (VP2) gene of the FPV strain currently prevalent in South Korea in a baculovirus expression system; VP2 protein was identified by an indirect immunofluorescence assay, electron microscopy (EM), Western blotting (WB), and a hemagglutination assay (HA). EM showed that the recombinant VP2 protein self-assembled to form virus-like particles. WB revealed that the recombinant VP2 was 65 kDa in size. The HA activity of the recombinant VP2 protein was very high at 1:215. A total of 143 cat serum samples were tested using FPV (HI-FPV test) and the recombinant VP2 protein (HI-VP2 test) as HI antigens. The sensitivity, specificity, and accuracy of the HI-VP2 test were 99.3%, 88.9%, and 99.3%, respectively, compared to the HI-FPV test. The HI-VP2 and HI-FPV results correlated significantly (r = 0.978). Thus, recombinant VP2 can substitute for intact FPV as the serological diagnostic reagent of the HI test for FPV.
Keywords
feline panleukopenia virus; VP2 protein; baculovirus expression system; hemagglutination inhibition test;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Stuetzer B, Hartmann K. Feline parvovirus infection and associated diseases. Vet J 2014;201:150-155.   DOI
2 Parrish CR. Pathogenesis of feline panleukopenia virus and canine parvovirus. Baillieres Clin Haematol 1995;8:57-71.   DOI
3 Yang S, Wang S, Feng H, Zeng L, Xia Z, Zhang R, Zou X, Wang C, Liu Q, Xia X. Isolation and characterization of feline panleukopenia virus from a diarrheic monkey. Vet Microbiol 2010;143:155-159.   DOI
4 Kim SG, Lee KI, Kim HJ, Park HM. Prevalence of feline panleukopenia virus in stray and household cats in Seoul, Korea. J Vet Clin 2013;30:333-338.
5 Kelly WR. An enteric disease of dogs resembling feline panleucopaenia. Aust Vet J 1978;54:593.   DOI
6 Allison AB, Organtini LJ, Zhang S, Hafenstein SL, Holmes EC, Parrish CR. Single mutations in the VP2 300 loop region of the three-fold spike of the carnivore parvovirus capsid can determine host range. J Virol 2015;90:753-767.   DOI
7 Su WJ, Shen WD, Li B, Wu Y, Gao G, Wang WB. A novel way to purify recombinant baculoviruses by using bacmid. Biosci Rep 2009;29:71-75.   DOI
8 Elia G, Desario C, Pezzoni G, Camero M, Brocchi E, Decaro N, Martella V, Buonavoglia C. Recombinant ELISA using baculovirus-expressed VP2 for detection of antibodies against canine parvovirus. J Virol Methods 2012;184:98-102.   DOI
9 Cheng N, Zhao Y, Han Q, Zhang W, Xi J, Yu Y, Wang H, Li G, Gao Y, Yang S, Liu W, Xia X. Development of a reverse genetics system for a feline panleukopenia virus. Virus Genes 2019; 55:95-103.   DOI
10 Jiao C, Zhang H, Liu W, Jin H, Liu D, Zhao J, Feng N, Zhang C, Shi J. Construction and immunogenicity of virus-like particles of feline parvovirus from the tiger. Viruses 2020;12:315.   DOI
11 Yang DK, Yoon SS, Byun JW, Lee KW, Oh YI, Song JY. Serological survey for canine parvovirus type 2a (CPV-2a) in the stray dogs in South Korea. J Bacteriol Virol 2010;40:77-81.   DOI
12 Dall'Ara P, Lauzi S, Filipe J, Caseri R, Beccaglia M, Desario C, Cavalli A, Aiudi GG, Buonavoglia C, Decaro N. Discrepancy between in-clinic and haemagglutination-inhibition tests in detecting maternally-derived antibodies against canine parvovirus in puppies. Front Vet Sci 2021;8:630809.   DOI
13 Jin H, Xia X, Liu B, Fu Y, Chen X, Wang H, Xia Z. High-yield production of canine parvovirus virus-like particles in a baculovirus expression system. Arch Virol 2016;161:705-710.   DOI
14 Shackelton LA, Hoelzer K, Parrish CR, Holmes EC. Comparative analysis reveals frequent recombination in the parvoviruses. J Gen Virol 2007;88(Pt 12):3294-3301.   DOI
15 Chang D, Liu Y, Chen Y, Hu X, Burov A, Puzyr A, Bondar V, Yao L. Study of the immunogenicity of the VP2 protein of canine parvovirus produced using an improved Baculovirus expression system. BMC Vet Res 2020;16:202.   DOI
16 Park JS, Choi BK, Vijayachandran LS, Ayyappan V, Chong CK, Lee KS, Kim SC, Choi CW. Immunodetection of canine parvovirus (CPV) in clinical samples by polyclonal antisera against CPV-VP2 protein expressed in Esherichia coli as an antigen. J Virol Methods 2007;146:281-287.   DOI
17 Chambers AC, Aksular M, Graves LP, Irons SL, Possee RD, King LA. Overview of the baculovirus expression system. Curr Protoc Protein Sci 2018 91:5.4.1-5.4.6.
18 Shin A, Lee M, Kim S, Kang SH. On-line capillary electrophoresis for enhanced detection sensitivity of feline panleukopenia virus. J Chromatogr B Analyt Technol Biomed Life Sci 2012;909:22-25.   DOI
19 Chapman MS, Rossmann MG. Structure, sequence, and function correlations among parvoviruses. Virology 1993;194: 491-508.   DOI
20 Molina A, Veramendi J, Hervas-Stubbs S. Induction of neutralizing antibodies by a tobacco chloroplast-derived vaccine based on a B cell epitope from canine parvovirus. Virology 2005;342:266-275.   DOI
21 Balboni A, Bassi F, De Arcangeli S, Zobba R, Dedola C, Alberti A, Battilani M. Molecular analysis of carnivore Protoparvovirus detected in white blood cells of naturally infected cats. BMC Vet Res 2018;14:41.   DOI
22 Chung HC, Kim SJ, Nguyen VG, Shin S, Kim JY, Lim SK, Park YH, Park B. New genotype classification and molecular characterization of canine and feline parvoviruses. J Vet Sci 2020;21:e43.   DOI
23 Abd-Eldaim M, Beall MJ, Kennedy MA. Detection of feline panleukopenia virus using a commercial ELISA for canine parvovirus. Vet Ther 2009;10:E1-E6.
24 Mira F, Purpari G, Lorusso E, Di Bella S, Gucciardi F, Desario C, Macaluso G, Decaro N, Guercio A. Introduction of Asian canine parvovirus in Europe through dog importation. Transbound Emerg Dis 2018;65:16-21.   DOI
25 Horiuchi M, Yamaguchi Y, Gojobori T, Mochizuki M, Nagasawa H, Toyoda Y, Ishiguro N, Shinagawa M. Differences in the evolutionary pattern of feline panleukopenia virus and canine parvovirus. Virology 1998;249:440-452.   DOI
26 Li G, Ji S, Zhai X, Zhang Y, Liu J, Zhu M, Zhou J, Su S. Evolutionary and genetic analysis of the VP2 gene of canine parvovirus. BMC Genomics 2017;18:534.   DOI