Babesia ovata: isolation from erythrocytes and development of an enzyme-linked immunosorbent assay for detection of antibodies.
A method of isolating Babesia ovata merozoites from infected erythrocytes and an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-B. ovata antibodies were developed. After B. 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 with B. ovata.
Shimizu
S
S
Laboratory of Protozoa, National Institute of Animal Health, Tsukuba Science City, Japan.
Shimura
K
K
Ito
S
S
Minami
T
T
D000818:Animals; D000913:Antibodies, Protozoan / Q000032:analysis*; D001403:Babesia / Q000276:immunology / Q000302:isolation & purification*; D001404:Babesiosis / Q000097:blood / Q000175:diagnosis* / Q000469:parasitology; D002417:Cattle; D002498:Centrifugation; D002499:Centrifugation, Density Gradient; D003168:Complement Fixation Tests; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000469:parasitology*; D005455:Fluorescent Antibody Technique; D012680:Sensitivity and Specificity
D000913:Antibodies, Protozoan