Echinococcus granulosus Protoscolex DM9 Protein Shows High Potential for Serodiagnosis of Alveolar Echinococcosis

Jeong-Geun Kim, Xiumin Han, Y. Kong
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Abstract

Alveolar echinococcosis (AE) caused by infection with E. multilocularis metacestode, represents one of the most fatal helminthic diseases. AE is principally manifested with infiltrative, proliferating hepatic mass, resembling primary hepatocellular carcinoma. Sometimes metastatic lesions are found in nearby or remote tissue. AE diagnosis largely depends on imaging studies, but atypical findings of imaging features frequently require differential diagnosis from other hepatic lesions. Serological tests may provide further evidence, while obtaining reliable AE materials is not easy. In this study, alternative antigens, specific to AE were identified by analyzing E. granulosus protoscolex proteins. An immunoblot analysis of E. granulosus protoscolex showed that a group of low-molecular-weight proteins in the range from 14 kDa to 16 kDa exhibited a sensitive and specific immune response to AE patient sera. Partial purification and proteomic analysis indicated that this protein group contained myosin, tubulin polymerization promoting protein, fatty-acid binding protein, uncharacterized DM9, heat shock protein 90 cochaperone tebp P-23, and antigen S. When the serological applicability of recombinant forms of these proteins was assessed using enzyme-linked immunosorbent assay, DM9 protein (rEgDM9) showed 90.1% sensitivity (73/81 sera tested) and 94.5% specificity (172/181 sera tested), respectively. rEgDM9 showed weak cross-reactions with patient sera from the transitional and chronic stages of cystic echinococcosis (3 to 5 stages). rEgDM9 would serve as a useful alternative antigen for serodiagnosis of both early- and advanced-stage AE cases.
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细粒棘球蚴原头节DM9蛋白在肺泡棘球蚴病的血清诊断中具有很高的潜力
肺泡棘球蚴病(AE)是由多房棘球蚴感染引起的最致命的寄生虫病之一。AE主要表现为浸润性、增生性肝脏肿块,类似原发性肝细胞癌。有时在附近或远处的组织中发现转移性病变。AE的诊断在很大程度上依赖于影像学检查,但影像学特征的非典型表现往往需要与其他肝脏病变鉴别诊断。血清学检测可提供进一步的证据,而获得可靠的声发射材料并不容易。在这项研究中,通过分析E. granulosus原头节蛋白,确定了AE特异性的替代抗原。免疫印迹分析表明,一组14 ~ 16 kDa范围内的低分子量蛋白对AE患者血清表现出敏感和特异性的免疫应答。部分纯化和蛋白质组学分析表明,该蛋白组含有肌球蛋白、微管蛋白聚合促进蛋白、脂肪酸结合蛋白、未表征的DM9、热休克蛋白90 cochaperone tebp P-23和抗原s。用酶联免疫吸附法评估这些蛋白重组形式的血清学适用性,DM9蛋白(rEgDM9)的敏感性为90.1%(73/81血清),特异性为94.5%(172/181血清)。rEgDM9与囊性包虫病过渡期和慢性期(3 ~ 5期)患者血清呈弱交叉反应。rEgDM9可作为早期和晚期AE病例血清诊断的有用替代抗原。
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