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微小巴贝斯虫一种新型分泌抗原1的鉴定与特性分析及其在酶联免疫吸附测定和免疫层析试验中的潜在应用评估

Identification and characterization of a novel secreted antigen 1 of Babesia microti and evaluation of its potential use in enzyme-linked immunosorbent assay and immunochromatographic test.

作者信息

Luo Yuzi, Jia Honglin, Terkawi M Alaa, Goo Youn-Kyoung, Kawano Suguru, Ooka Hideo, Li Yan, Yu Longzheng, Cao Shinuo, Yamagishi Junya, Fujisaki Kozo, Nishikawa Yoshifumi, Saito-Ito Atsuko, Igarashi Ikuo, Xuan Xuenan

机构信息

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.

出版信息

Parasitol Int. 2011 Jun;60(2):119-25. doi: 10.1016/j.parint.2010.11.001. Epub 2010 Nov 8.

Abstract

Here, we identified a novel secreted antigen designated as Babesia microti secreted antigen 1 (BmSA1) by immunoscreening a B. microti cDNA expression library using the sera from hamsters immunized with plasma, putatively containing secreted antigens, from B. microti-infected hamsters. Antibodies raised in mice immunized with recombinant BmSA1 (rBmSA1) recognized a native 33-kDa parasite protein. An enzyme-linked immunosorbent assay (ELISA) of rBmSA1 detected specific antibodies as early as 6 and 4 days post-infection in sera from a hamster experimentally infected with B. microti Gray strain (US type) and a mouse experimentally infected with B. microti Munich strain (rodent isolate), respectively. Moreover, a rapid immunochromatographic test (ICT) using rBmSA1 detected specific antibodies in a hamster experimentally infected with B. microti from day 6 to at least day 270 post-infection, which was quite consistent with the results of the ELISA. In addition, analysis of the sera involved in the first case of human babesiosis in Japan (Kobe type) showed that specific antibodies were detectable in the patient and the positive donor by ELISA using rBmSA1, and the ICT result was identical to the ELISA data. Taken together, these results indicated that BmSA1 could be a promising and universal target for developing both ELISA and ICT for the serodiagnosis of human babesiosis and for an epidemiological survey of its rodent reservoir.

摘要

在此,我们通过使用来自感染微小巴贝斯虫的仓鼠血浆(推测含有分泌抗原)免疫的仓鼠血清对微小巴贝斯虫cDNA表达文库进行免疫筛选,鉴定出一种名为微小巴贝斯虫分泌抗原1(BmSA1)的新型分泌抗原。用重组BmSA1(rBmSA1)免疫的小鼠产生的抗体识别出一种天然的33 kDa寄生虫蛋白。rBmSA1的酶联免疫吸附测定(ELISA)分别在实验感染微小巴贝斯虫格雷株(美国型)的仓鼠血清和实验感染微小巴贝斯虫慕尼黑株(啮齿动物分离株)的小鼠血清中,感染后第6天和第4天就检测到了特异性抗体。此外,使用rBmSA1的快速免疫层析试验(ICT)在实验感染微小巴贝斯虫的仓鼠中,从感染后第6天到至少第270天检测到了特异性抗体,这与ELISA结果相当一致。另外,对日本首例人类巴贝斯虫病(神户型)相关血清的分析表明,使用rBmSA1的ELISA在患者和阳性供体中可检测到特异性抗体,ICT结果与ELISA数据一致。综上所述,这些结果表明BmSA1可能是开发用于人类巴贝斯虫病血清诊断的ELISA和ICT以及对其啮齿动物宿主进行流行病学调查的有前景的通用靶点。

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