Parasitology Research

, Volume 78, Issue 8, pp 684–688 | Cite as

Babesia ovata: isolation from erythrocytes and development of an enzyme-linked immunosorbent assay for detection of antibodies

  • S. Shimizu
  • K. Shimura
  • S. Ito
  • T. Minami
Original Investigations
Accepted:

Abstract

A method of isolatingBabesia ovata merozoites from infected erythrocytes and an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-B. ovata antibodies were developed. AfterB. ovata-infected erythrocytes had been lysed using the nitrogen cavitation method, the merozoites were separated from erythrocyte components by differential centrifugation and density-gradient centrifugation. The light microscopic examination showed that the purified merozoites were morphologically intact and contained few contaminants. Sodium dodecyl sulfate-polyacrylamide electrophoretic (SDS-PAGE) analysis revealed that the merozoite fraction contained very little contamination with erythrocyte components. The merozoites thus obtained were sonicated and treated with Triton X-100 and then used as an antigen to measure anti-B. ovata antibodies in ELISA. The ELISA was more sensitive in detecting anti-B. ovata antibodies than was either the indirect fluorescent antibody test or the complement fixation test on sera from cattle infected withB. ovata.

Keywords

Dodecyl Fixation Test Antibody Test Differential Centrifugation Fluorescent Antibody 
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.
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Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • S. Shimizu
    • 1
  • K. Shimura
    • 1
  • S. Ito
    • 1
  • T. Minami
    • 1
  1. 1.Laboratory of Protozoa, First divisionNational Institue of Animal HealthTsukuba Science CityJapan

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