CAJ | 학술논문

吉氏巴贝斯虫BgTRAP蛋白是一个重要的诊断抗原候选分子。为建立一种实用的犬吉氏巴贝斯虫血清学诊断方法，本研究选取BgTRAP C-末端跨膜区前，包含TSP功能区和抗原区的411个氨基酸的编码基因片段，重组表达了一个可溶性截短型BgTRAP抗原，解决了完整蛋白重组表达纯化的困难。免疫荧光试验表明，截短型抗原具有良好的抗原性。纯化的截短型抗原作为酶联免疫试验诊断抗原，可清晰地区分阴性及阳性犬血清，并与其他病原感染无交叉反应，具有良好的特异性。犬感染吉氏巴贝斯虫系列血清检测表明，该抗原可检测早期感染（4 d）和感染200 d以后的样本。结果提示，重组BgTRAP截短型抗原可作为一种诊断制剂检测犬吉氏巴贝斯虫抗体。
길씨파패사충BgTRAP단백시일개중요적진단항원후선분자。위건립일충실용적견길씨파패사충혈청학진단방법，본연구선취BgTRAP C-말단과막구전，포함TSP공능구화항원구적411개안기산적편마기인편단，중조표체료일개가용성절단형BgTRAP항원，해결료완정단백중조표체순화적곤난。면역형광시험표명，절단형항원구유량호적항원성。순화적절단형항원작위매련면역시험진단항원，가청석지구분음성급양성견혈청，병여기타병원감염무교차반응，구유량호적특이성。견감염길씨파패사충계렬혈청검측표명，해항원가검측조기감염（4 d）화감염200 d이후적양본。결과제시，중조BgTRAP절단형항원가작위일충진단제제검측견길씨파패사충항체。
The BgTRAP protein of Babesia gibsoni is an important candidate for the development of a diagnostic reagent for canine babesiosis. In order to develop a diagnostic method for practical use, the gene fragment encoding truncated BgTRAP that covered TSP region and antigenic area ahead of transmembrane region was cloned and expressed in E.coli in soluble form. The recombinant truncated BgTRAP protein showed strong antigenicity in indirect fluorescent-antibody test and ELISA. The ELISA method using truncated BgTRAP protein was able to differentiate B. gibsoni-infected dog serum and non-infected dog serum. In dogs experimentally infected with B. gibsoni, ELISA method also detected antibody response as early as 4 days, or as late as 200 days post infection. These results demonstrated that the recombinant truncated BgTRAP protein might be a useful diagnostic reagent for detection of B.gibsoni antibodies in dogs.