Cloning of a truncated Babesia equi gene encoding an 82-kilodalton protein and its potential use in an enzyme-linked immunosorbent assay.

Journal of Clinical Microbiology
Haruyuki HirataIkuo Igarashi

Abstract

To isolate Babesia equi genes encoding immunodominant proteins, a cDNA expression library prepared from B. equi mRNA was immunoscreened with B. equi-infected horse serum. Eighteen positive cDNA clones were obtained, and the clone that showed the strongest immunoreactivity, designated Be82, was further characterized. The Be82 gene consisted of 1,953 bp and contained a partial open reading frame lacking the 5'-terminal sequence. As shown by Western blot analyses, immune sera from mice intraperitoneally injected with the Be82 gene product recognized the 82- and 52-kDa proteins of B. equi but not those of Babesia caballi. The glutathione S-transferase fusion protein expressed in Escherichia coli that was purified and used as the antigen in the enzyme-linked immunosorbent assay reacted specifically with B. equi-infected horse sera. These results suggest that the Be82 gene product is a potential diagnostic antigen candidate in the detection of B. equi infection in horses that will be useful both in the performance of epidemiological studies and in the granting of quarantine passes.

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Citations

Dec 25, 2003·Antimicrobial Agents and Chemotherapy·Sabine BorkIkuo Igarashi
Jul 13, 2012·The Veterinary Journal·Ignacio García-BocanegraAntonio Arenas
Feb 27, 2010·Veterinary Parasitology·Pierre-Hugues PitelGuillaume Fortier
Apr 2, 2013·Molecular and Biochemical Parasitology·Marta G SilvaDonald P Knowles
Sep 19, 2006·Molecular and Biochemical Parasitology·Hiromi IkadaiTakashi Oyamada
Mar 29, 2011·Revista brasileira de parasitologia veterinária = Brazilian journal of veterinary parasitology : Órgão Oficial do Colégio Brasileiro de Parasitologia Veterinária·Cristiane Divan BaldaniRosangela Zacarias Machado
May 10, 2006·Clinical and Vaccine Immunology : CVI·Xiaohong HuangIkuo Igarashi
Feb 9, 2005·Clinical and Diagnostic Laboratory Immunology·Haruyuki HirataIkuo Igarashi

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