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The aim of this study was to identify early specific diagnostic antigens in<jats:italic>T. spiralis<\/jats:italic>ES proteins with 30\u201340\u2009kDa. The ES proteins were analyzed by two-dimensional electrophoresis (2-DE), and a total of approximately 150 proteins spots were detected with isoelectric point (pI) varying from 4 to 7 and molecular weight from 14 to 66\u2009kDa. When probed with sera from infected mice at 18 days postinfection, ten protein spots with molecular weight of 30\u201340\u2009kDa were recognized and identified by MALDI-TOF\/TOF-MS. All of ten spots were successfully identified and characterized to correlate with five different proteins, including two potential serine proteases, one antigen targeted by protective antibodies, one deoxyribonuclease (DNase) II, and one conserved hypothetical protein. 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