Advertisement

Parasitology Research

, Volume 105, Issue 1, pp 249–254 | Cite as

Evaluation of Babesia bigemina 200 kDa recombinant antigen in enzyme-linked immunosorbent assay

  • Khukhuu Altangerel
  • Andy Alhassan
  • Hiroshi Iseki
  • Thillaiampalam Sivakumar
  • Damdinsuren Boldbaatar
  • Naoaki Yokoyama
  • Ikuo IgarashiEmail author
Original Paper

Abstract

A truncated fragment of the gene encoding the 200-kDa protein (P200) of Babesia bigemina was cloned into a plasmid vector, pGEX-4 T-1 and expressed in Escherichia coli as a glutathione-S-transferase fused protein. An indirect enzyme-linked immunosorbent assay (ELISA) using the rp200/CT detected specific antibodies in cattle experimentally infected with B. bigemina. Furthermore, the antigen did not cross-react with antibodies to Babesia bovis, a closely related Babesia parasite indicating that rp200/CT is a specific antigen for the diagnosis of B. bigemina infection. Additionally, ELISA using p200/CT and polymerase chain reaction were conducted on serum and corresponding DNA samples obtained from field cattle to evaluate the diagnostic utility of the p200/CT antigen. Results from the current study suggest that p200/CT ELISA is a sensitive and specific method for improved serodiagnosis of B. bigemina infection.

Keywords

Polymerase Chain Reaction Babesia Babesiosis Infected Cattle Ampicillin Sodium 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This study was supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science, a Program for Promotion of Basic Research Activities for Innovative Biosciences, and grants from the twenty-first century COE Program and the Program of Founding Research Centers for Emerging and Reemerging Infectious Diseases, the Ministry of Education, Culture, Sports, Science, and Technology of Japan, and the Japan International Cooperation Agency.

References

  1. Aguilar-Delfin MJ, Homer PJ, Wettstein PJ, Persing DH (2001) Innate resistance to Babesia infection is influenced by genetic background and gender. Infect Immun 69:7955–7958CrossRefPubMedGoogle Scholar
  2. Aguilar-Delfin PJ, Wettstein PJ, Persing DH (2003) Resistance to acute babesiosis is associated with interleukin-12- and gamma interferon-mediated responses and requires macrophages and natural killer cells. Infect Immun 71:2002–2008CrossRefPubMedGoogle Scholar
  3. Akinboade OA, Akinboade CY (1985) The effect of Babesia bigemina infections caused by cattle ticks on Nigerian economy. Rev Elev Med Vet Pays Trop 38:250–252PubMedGoogle Scholar
  4. Araujo FR, Madruga CR, Leal CR, Schenk MA, Kessler RH, Marques AP, Lemaire DC (1998) Comparison between enzyme-linked immunosorbent assay, indirect fluorescent antibody and rapid conglutination tests in detecting antibodies against Babesia bovis. Vet Parasitol 74:101–108CrossRefPubMedGoogle Scholar
  5. Barnett SF (1974) Economical aspects of protozoal tick-borne diseases in livestock in parts of the world other than Britain. Bull Off int Epiz 8:183–196Google Scholar
  6. Boonchit S, Xuan X, Yokoyama N, Goff WL, Wagner G, Igarashi I (2002) Evaluation of an enzyme-linked immunosorbent assay with recombinant rhoptry-associated protein 1 antigen against Babesia bovis for the detection of specific antibodies in cattle. J Clin Microbiol 40:3771–3775CrossRefPubMedGoogle Scholar
  7. Boonchit S, Alhassan A, Chan B, Xuan X, Yokoyama N, Ooshiro M, Goff WL, Waghela SD, Wagner G, Igarashi I (2006) Expression of C-terminal truncated and full-length Babesia bigemina rhoptry-associated protein 1 and their potential use in enzyme-linked immunosorbent assay. Vet Parasitol 137:28–35CrossRefPubMedGoogle Scholar
  8. Böse R, Jorgensen WK, Dalgliesh RJ, Friendhoff KT, de Vos AJ (1995) Current state and future trends in the diagnosis of babesiosis. Vet Parasitol 57:61–74CrossRefPubMedGoogle Scholar
  9. Brown WC, Norimine J, Knowles DP, Goff WL (2006) Immune control of Babesia bovis infection. Vet Parasitol 138:75–87CrossRefPubMedGoogle Scholar
  10. Dalrymple BP, Jorgensen WK, de Vos AJ, Wright IG (1992) Analysis of the composition of samples of Babesia bovis and the influence of different environmental conditions on genetically distinct subpopulations. Int J Parasitol 22:731–737CrossRefPubMedGoogle Scholar
  11. De Vos AJ, Bock RE (2000) Vaccination against bovine babesiosis. Ann NY Acad Sci 916:540–545PubMedCrossRefGoogle Scholar
  12. El-Ghaysh A, Sundquist B, Christensson DA, Hilali M, Nassar AM (1996) Observations on the use of ELISA for detection of Babesia bigemina specific antibodies. Vet Parasitol 62:51–61CrossRefPubMedGoogle Scholar
  13. Goff WL, Johnson WC, Molloy JB, Jorgensen WK, Waldron SJ, Figueroa JV, Matthee O, Adams DS, McGuire TC, Pino I, Mosqueda J, Palmer GH, Suarez CE, Knowles DP, McElwain TF (2008) Validation of a competitive ELISA for detection of Babesia bigemina antibodies in cattle. Clin Vaccine Immunol 15:1316–1321CrossRefPubMedGoogle Scholar
  14. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227:680–685CrossRefGoogle Scholar
  15. Mbati PA, Hlatshwayo M, Mtshali MS, Mogaswane KR, de Waal TD, Dipeolu OO (2002) Ticks and tick-borne diseases of livestock belonging to resource-poor farmers in the eastern Free State of South Africa. Exp Appl Acarol 28:217–224CrossRefPubMedGoogle Scholar
  16. McCosker PJ (1981) The global importance of babesiosis, pp. 1–24. In: Ristic M, Kreier JP (eds) babesiosis. Academic Press, New YorkGoogle Scholar
  17. Molloy JB, Bowles PM, Jeston PJ, Bruyeres AG, Bowden JM, Bock RE, Jorgensen WK, Blight GW, Dalgliesh RJ (1998) Development of an enzyme-linked immunosorbent assay for detection of antibodies to Babesia bigemina in cattle. Parasitol Res 84:651–656CrossRefPubMedGoogle Scholar
  18. Morzaria S, Katende J, Kairo A, Nene V, Musoke A (1992) New methods for the diagnosis of Babesia bigemina infection. Mem Inst Oswaldo Cruz 87:201–205CrossRefPubMedGoogle Scholar
  19. Ravindran R, Mishra AK, Rao JR (2007) Slide enzyme-linked immunosorbent assay for the diagnosis of Babesia bigemina infection in bovines. Vet Res Commun 31:999–1004CrossRefPubMedGoogle Scholar
  20. Solari MA, Nari A, Cardozo H (1992) Impact of Babesia bovis and Babesia bigemina on the production of beef cattle in Uruguay. Mem Inst Oswaldo Cruz 87:143–149CrossRefPubMedGoogle Scholar
  21. Stegeman JR, Birkenheuer AJ, Kruger JM, Breitschwerdt EB (2003) Transfusion associated Babesia gibsoni infection in a dog. J Am Vet Med Assoc 222:959–963CrossRefPubMedGoogle Scholar
  22. Tebele N, Skilton RA, Katende J, Wells CW, Nene V, McElwain T, Morzaria SP, Musoke AJ (2000) Cloning, characterization, and expression of a 200-kilodalton diagnostic antigen of Babesia bigemina. J Clin Microbiol 38:2240–2247PubMedGoogle Scholar
  23. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354CrossRefPubMedGoogle Scholar
  24. Vega CA, Buening GM, Green TJ, Carson CA (1985) In vitro cultivation of Babesia bigemina. Am J Vet Res 46:416–420PubMedGoogle Scholar
  25. Weiland G, Reiter I (1988) Methods for serological response to babesia. In: Ristic M (ed) Babesiosis of domestic animal and man. CRC Press, Boca Raton, Fla, pp 143–158Google Scholar
  26. Wright IG, Goodger BV, Leatch G, Aylward JH, Rode-Bramanis K, Waltisbuhl DJ (1987) Protection of Babesia bigemina-immune animals against subsequent challenge with virulent Babesia bovis. Infect Immun 55:364–368PubMedGoogle Scholar
  27. Xuan X, Larsen A, Ikadai H, Tanaka T, Igarashi I, Nagasawa H, Fujisaki K, Toyoda Y, Suzuki N, Mikami T (2001) Expression of Babesia equi merozoite antigen 1 in insect cells by recombinant baculovirus and evaluation of its diagnostic potential in an enzyme-linked immunosorbent assay. J Clin Microbiol 39:705–709CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Khukhuu Altangerel
    • 1
  • Andy Alhassan
    • 1
  • Hiroshi Iseki
    • 1
  • Thillaiampalam Sivakumar
    • 1
  • Damdinsuren Boldbaatar
    • 2
  • Naoaki Yokoyama
    • 1
  • Ikuo Igarashi
    • 1
    Email author
  1. 1.National Research Center for Protozoan DiseasesObihiro University of Agriculture and Veterinary MedicineObihiroJapan
  2. 2.Laboratory of Emerging Infectious Diseases, Department of Frontier Veterinary Medicine, Faculty of AgricultureKagoshima UniversityKorimotoJapan

Personalised recommendations