A cDNA expression library prepared from
Babesia gibsoni (
B. gibsoni) merozoite mRNA was screened with a
B. gibsoni-infected dog serum. A cDNA clone encoding 30 kDa protein was cloned and designated P30 gene. The complete nucleotide sequence of the P30 gene had 792 bp. Computer analysis suggested that the sequence of the P30 gene contained an open reading frame of 600 bp with a coding capacity of approximately 23 kDa. The native P30 protein of
B. gibsoni with molecular mass of 30 kDa was identified by Westem blotting using anti-recombinant P30 mouse serum. The recombinant P30 protein expressed in
Escherichia coli was used for antigen in an enzyme-linked immunosorbent assay (ELISA). The ELISA was able to detect the
B. gibsoni infection and did not show any cross reactivity to
B. canis-infected dog sera and normal dog sera. Furthermore, the antibody response against the recombinant P30 was maintained in dog experimentally infected with
B. gibsoni even after the dog became the chronic stage of infection. These results demonstrated that the recombinant P30 might be a useful diagnostic reagent for detection of antibodies to
B. gibsoni in dogs.
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