A novel recombinant antigen for immunodiagnosis of human cystic echinococcosis
A novel recombinant antigen for immunodiagnosis of human cystic echinococcosis
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DOI：
10.1086/379976
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发表时间：
2003-12-15
影响因子：
6.4
通讯作者：
McManus, DP
中科院分区：
文献类型：
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作者：
Li, J;Zhang, WB;McManus, DP
用细粒棘球蚴感染小鼠的血清样品池筛选由羊包虫囊原头节幼虫RNA构建的cDNA文库。一个免疫反应性克隆，命名为EpC 1，被证明编码的蛋白质，76个残基。将互补DNA（cDNA）片段亚克隆到表达载体pET-41 b（+）中，并在大肠杆菌中表达所得重组EpC 1谷胱甘肽S-转移酶（GST）融合蛋白（rEpC 1-GST），并针对GST标签进行亲和纯化。免疫球蛋白G是针对rEpC 1-GST产生的优势抗体同种型。用免疫球蛋白G免疫印迹法检测896例人血清标本中融合蛋白的诊断敏感性和特异性; 324份来自囊型包虫病（CE）患者的血清样本，172份来自神经囊尾蚴病患者的血清样本，89份来自泡型包虫病患者的血清样本，以及241份来自其他感染或临床表现患者的血清样本，以及70例确认阴性对照受试者，总灵敏度为92.2%，总特异性为95.6%。考虑到测试的大量血清样本，rEpC 1-GST融合蛋白诊断CE的灵敏度和特异性水平是前所未有的。
A pool of serum samples from mice infected with oncospheres (eggs) of Echinococcus granulosus was used to screen a cDNA library constructed with RNA extracted from protoscolex larvae from sheep hydatid cysts. One immunoreactive clone, designated EpC1, was shown to encode a protein, of 76 residues. The complementary DNA (cDNA) fragment was subcloned into an expression vector, pET-41b(+), and the resulting recombinant EpC1 glutathione S-transferase (GST) fusion protein (rEpC1-GST) was expressed in Escherichia coli and was affinity purified against the GST tag. Immunoglubulin G was the dominant antibody isotypes generated against rEpC1-GST. A total of 896 human serum samples were used to evaluate the diagnostic sensitivity and specificity of the fusion protein by immunoglobulin G immunoblotting; 324 serum samples from patients with cystic echinococcosis (CE), 172 from patients with neurocysticercosis, 89 from patients with alveolar echinococcosis, and 241 from patients with other infections or clinical presentations, as well as 70 from confirmed-negative control subjects, yielded an overall sensitivity of 92.2% and an overall specificity of 95.6%. The combined levels of sensitivity and specificity achieved with the rEpC1-GST fusion protein for diagnosis of CE are unprecedented, taking into account the large panel of serum samples that were tested.