Molecular identification and antigenic characterization of a merozoite surface antigen and a secreted antigen of Babesia canis (BcMSA1 and BcSA1).
Molecular identification and antigenic characterization of a merozoite surface antigen and a secreted antigen of Babesia canis (BcMSA1 and BcSA1).
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DOI：
10.1186/s13071-016-1518-1
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发表时间：
2016-05-03
影响因子：
3.2
通讯作者：
Xuan X
中科院分区：
文献类型：
--
作者：
Zhou M;Cao S;Luo Y;Liu M;Wang G;Moumouni PF;Jirapattharasate C;Iguchi A;Vudriko P;Terkawi MA;Löwenstein M;Kern A;Nishikawa Y;Suzuki H;Igarashi I;Xuan X
犬巴贝虫是一种顶复形蜱传播的血原虫，在欧洲和西亚引起犬巴贝虫病。尽管它的重要性，没有已知的快速诊断试剂盒检测B。犬感染。本研究鉴定了两种新的B抗原。建立了快速、特异、敏感的犬B型B血清学诊断技术。犬感染。从B的mRNA中构建互补DNA（cDNA）表达文库。犬，并使用B进行免疫筛选。犬感染的狗血清鉴定了编码裂殖子表面抗原和分泌抗原蛋白的cDNA，并分别命名为BcMSA 1和BcSA 1。纯化表达的重组BcMSA 1和BcSA 1（rBcMSA 1和rBcSA 1），免疫小鼠制备抗血清。天然蛋白的特征在于通过Western印迹分析和免疫荧光。建立了基于rBcMSA 1或rBcSA 1的间接酶联免疫吸附试验（iELISA）和快速免疫层析试验（ICT），并对两个B患者连续血浆样本中的特异性抗体进行检测。感染犬的狗通过Western印迹分析，针对rBcMSA 1和rBcSA 1的抗血清分别识别39 kDa和44 kDa的天然蛋白。此外，免疫荧光和共聚焦显微镜观察显示，BcMSA 1被发现在寄生虫的表面。BcSA 1定位于裂殖子的基质中。以rBcMSA 1或rBcSA 1为基础的ELISA和ICT可检测到两个B的连续血浆样品中的特异性抗体。感染犬的狗它们与从实验性感染密切相关寄生虫的狗中采集的血清样品没有交叉反应。综上所述，目前的结果表明，rBcMSA 1和rBcSA 1是有前途的血清学诊断抗原，用于发展iELISA和ICT检测B。犬感染。据我们所知，这项研究是第一个报告BcMSA 1和BcSA 1作为潜在的抗原蛋白的血清诊断B。犬感染。
Babesia canis is an apicomplexan tick-transmitted hemoprotozoan responsible for causing canine babesiosis in Europe and west Asia. Despite its importance, there is no known rapid diagnostic kit detection of B. canis infection in dogs. The present study identified two novel antigens of B. canis and used the recombinant antigens to establish a rapid, specific and sensitive serodiagnostic technique for detection of B. canis infection. A complementary DNA (cDNA) expression library was constructed from the mRNA of B. canis and immunoscreened using B. canis-infected dog sera. The cDNAs encoding a merozoite surface antigen and a secreted antigen protein were identified and designated as BcMSA1 and BcSA1, respectively. The recombinant BcMSA1 and BcSA1 (rBcMSA1 and rBcSA1) expressed in Escherichia coli were purified and injected into mice for production of anti-sera. The native proteins were characterized by Western blot analysis and immunofluorescence. Furthermore, indirect enzyme-linked immunosorbent assays (iELISA) and rapid immunochromatographic tests (ICT) based on rBcMSA1 or rBcSA1 were established and evaluated to test specific antibodies in consecutive plasma samples from two B. canis-infected dogs. Antiserum raised against rBcMSA1 and rBcSA1 recognized the 39 kDa and 44 kDa native proteins by Western blot analysis, respectively. In addition, immunofluorescence and confocal microscopic observations revealed that BcMSA1 was found on the surface of parasites. However, BcSA1 localized in the matrix of the merozoites. The ELISA and ICT based on rBcMSA1 or rBcSA1 could detect specific antibodies in consecutive plasma samples from two B. canis-infected dogs. They showed no cross-reactions against the serum samples collected from dogs experimentally infected with closely related parasites. Taken together, the current results indicated that the rBcMSA1 and rBcSA1 are promising serodiagnostic antigens for developing iELISA and ICT to detect B. canis infection. To our knowledge, this study is the first to report BcMSA1 and BcSA1 as potential antigenic proteins for serodiagnosis of B. canis infection in dogs.