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Title: DEVELOPMENT OF A COMPETITIVE ELISA TEST FOR BABESIA BOVIS INFECTIONS BASED ON MEROZOITE SURFACE ANTIGEN-2C

Author
item DOMINGUEZ, M - INTA, ARGENTINA
item ZABAL, O - INTA, ARGENTINA
item WILKOWSKY, S - INTA, ARGENTINA
item RODRIGUEZ, A - INTA, ARGENTINA
item ASENZO, G - INTA, ARGENTINA
item FARBER, M - INTA, ARGENTINA
item ECHAIDE, I - INTA, ARGENTINA
item ECHAIDE, S - INTA, ARGENTINA
item Suarez, Carlos
item FLORIN-CHRISTENSEN, M - INTA, ARGENTINA

Submitted to: Biocell
Publication Type: Abstract Only
Publication Acceptance Date: 11/1/2003
Publication Date: 11/1/2003
Citation: Dominguez, M., Zabal, O., Wilkowsky, S., Rodriguez, A., Asenzo, G., Farber, M., Echaide, I., Echaide, S.T., Suarez, C.E., Florin-Christensen, M. 2003. Development of a competitive elisa test for babesia bovis infections based on merozoite surface antigen-2c [abstract]. Biocell. 27:1-172.

Interpretive Summary: Previous studies have shown that Babesia bovis merozoite surface antigen-2c (MSA-2c) has a high degree of genetic and antigenic conservation among geographically distant strains. This antigen thus appears as an adequate diagnostic candidate for bovine babesiosis. We have produced a set of monoclonal antibodies against recombinant MSA-2c (rMSA-2c) from the Argentine R1A strain, expressed in a prokaryotic system. One of them, MAb H9P2C2, was shown to react in Western blots with a parasite protein of the expected MSA-2c size. MAb H9P2C2 and rMSA- 2c were employed to develop a competitive ELISA test. All tested B. bovis positive sera from experimental and natural infections were detected by the new cELISA test, while no false positives were observed with negative sera. These results indicate that this cELISA could be an adequate diagnostic tool for bovine babesiosis. Standardization of the test conditions and validation with a larger number of samples are currently under progress.

Technical Abstract: Previous studies have shown that Babesia bovis merozoite surface antigen-2c (MSA-2c) has a high degree of genetic and antigenic conservation among geographically distant strains. This antigen thus appears as an adequate diagnostic candidate for bovine babesiosis. We have produced a set of monoclonal antibodies against recombinant MSA-2c (rMSA-2c) from the Argentine R1A strain, expressed in a prokaryotic system. One of them, MAb H9P2C2, was shown to react in Western blots with a parasite protein of the expected MSA-2c size. MAb H9P2C2 and rMSA- 2c were employed to develop a competitive ELISA test. This test essentially consisted of binding of rMSA-2c to Immulon 2HB plates, blocking, incubations with a) serial dilutions of different sera from control or B. bovis-naturally or experimentally infected bovines, b) MAb H9P2C2, c) peroxidase-conjugated anti-mouse IgG and d) OPD-H2O2 colorimetric substrate, and optical density readings at 490nm. All tested B. bovis positive sera from experimental and natural infections significantly reduced binding of MAb H9P2C2 to rMSA-2c, while no reduction was observed with negative sera. These results indicate that this cELISA could be an adequate diagnostic tool for bovine babesiosis. Standardization of the test conditions and validation with a larger number of samples are currently under progress.