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首页> 外文期刊>Parasitology >Molecular characterization of a novel 32-kDa merozoite antigen of Babesia gibsoni with a better diagnostic performance by enzyme-linked immunosorbent assay.
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Molecular characterization of a novel 32-kDa merozoite antigen of Babesia gibsoni with a better diagnostic performance by enzyme-linked immunosorbent assay.

机译：通过酶联免疫吸附测定法对长柄巴贝斯球菌的新型32 kDa裂殖子抗原进行了分子表征，具有更好的诊断性能。

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摘要

We cloned and expressed a novel gene encoding a 32-kDa merozoite protein of Babesia gibsoni (BgP32). The length of nucleotide sequence of the cDNA was 1464 bp with an open reading frame of 969 bp. The truncated recombinant BgP32 (rBgP32) without a signal peptide and C-terminal hydrophobic sequence was expressed in Escherichia coli as a soluble glutathione-S-transferase (GST) fusion protein. Western blotting demonstrated that the native protein was 32-kDa, consistent with molecular weight of the predicted mature polypeptide. Enzyme-linked immunosorbent assay (ELISA) using rBgP32 detected specific antibodies from 8 days to 541 days post-infection in the sequential sera from a dog experimentally infected with B. gibsoni. Moreover, the antigen did not cross-react with B. canis subspecies and closely related protozoan parasites, indicating that rBgP32 is a specific diagnostic antigen. Analysis of 47 sera taken from dogs with anaemic signs revealed that rBgP32 detected a higher proportion of B. gibsoni seropositive samples (77%) than its previously identified rBgP50 (68%) homologue. These results indicate that the BgP32 is a novel immunodominant antigen of B. gibsoni, and rBgP32 might be useful for diagnosis of B. gibsoni infection.
机译：我们克隆并表达了一种新的基因，该基因编码巴比斯犬（Babesia gibsoni）（BgP32）的32 kDa裂殖子蛋白。 cDNA的核苷酸序列的长度为1464bp，开放阅读框为969bp。不含信号肽和C端疏水序列的截短重组BgP32（rBgP32）在大肠杆菌中以可溶性谷胱甘肽S-转移酶（GST）融合蛋白表达。蛋白质印迹证实天然蛋白质为32-kDa，与预测的成熟多肽的分子量一致。使用rBgP32的酶联免疫吸附测定（ELISA）在感染后8天至541天的连续血清中检测到了来自实验感染了吉布森牛的狗的特异性抗体。而且，该抗原没有与犬双歧杆菌亚种和紧密相关的原生动物寄生虫交叉反应，表明rBgP32是一种特异性诊断抗原。对从具有贫血迹象的狗中采集的47份血清进行的分析表明，rBgP32检测到的吉卜赛氏菌血清阳性样本比例（其先前鉴定的rBgP50（68％）更高）。这些结果表明，BgP32是一种长柄双歧杆菌的新型免疫优势抗原，而rBgP32可能对诊断长柄双歧杆菌感染有用。

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