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首页> 外文期刊>Journal of Clinical Microbiology >Alveolar Echinococcosis: Characterization of Diagnostic Antigen Em18 and Serological Evaluation of Recombinant Em18
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Alveolar Echinococcosis: Characterization of Diagnostic Antigen Em18 and Serological Evaluation of Recombinant Em18

机译:肺泡棘球cc病:诊断抗原Em18的表征和重组Em18的血清学评估

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The Echinococcus multilocularis protein Em18 is one of the most promising antigens for use in serodiagnosis of alveolar echinococcosis in human patients. Here we identify an antigenic relationship between Em18 and a 65-kDa immunodominant E. multilocularis surface protein previously identified as either EM10 or EmII/3. The NH2-terminal sequence of native Em18 was determined, revealing it to be a fragment of EM10. Experiments were undertaken to investigate the effect of proteinase inhibitors on the degradation of EM10 in crude extracts of E. multilocularis protoscoleces. Em18 was found to be the product of degradation of EM10 by cysteine proteinase. A recombinant Em18 (RecEm18, derived from 349K to 508K of EM10) was successfully expressed by using Escherichia coli expression system and then evaluated for use in serodiagnosis of alveolar echinococcosis. RecEm18 was recognized by 27 (87.1%) and 28 (90.3%) of 31 serum samples from clinically and/or pathologically confirmed alveolar echinococcosis patients by enzyme-linked immunosorbent assay and immunoblotting, respectively. Of 33 serum samples from cystic echinococcosis patients, 1 was recorded as having a weak positive reaction to RecEm18; however, none of the serum samples which were tested from neurocysticercosis patients (n = 10) or healthy people (n = 15) showed positive reactions. RecEm18 has the potential for use in the differential serodiagnosis of alveolar echinococcosis.
机译:多眼棘球球菌蛋白Em18是最有前途的抗原,可用于人类患者肺泡棘球菌病的血清学诊断。在这里,我们确定了Em18和65 kDa免疫优势 E之间的抗原关系。以前鉴定为EM10或EmII / 3的多眼表面蛋白。确定了天然Em18的NH 2 -末端序列,表明它是EM10的片段。进行实验以研究蛋白酶抑制剂对 E粗提物中EM10降解的影响。多眼结膜原癌。发现Em18是半胱氨酸蛋白酶降解EM10的产物。使用大肠杆菌表达系统成功表达了重组Em18(RecEm18,从EM10的 349 K衍生到 508 K),并对其进行了评估用于肺泡棘球菌病的血清诊断。通过酶联免疫吸附测定和免疫印迹,分别从临床和/或病理证实的肺泡棘球虫病患者的31个血清样本中分别识别了RecEm18(占27.(87.1%)和28(90.3%))。在来自囊性棘球co虫病患者的33份血清样品中,有1份对RecEm18的阳性反应较弱;但是,神经囊尾osis病患者( n = 10)或健康人( n = 15)检测的血清样本均未显示阳性反应。 RecEm18具有用于肺泡棘球菌病的血清学鉴别诊断的潜力。

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