使用重组抗原来进行华支睾吸虫病的免疫诊断。
Immunodiagnosis of clonorchiasis using a recombinant antigen.
作者信息
Yong T S, Yang H J, Park S J, Kim Y K, Lee D H, Lee S M
机构信息
Department of Parasitology, Yonsei University College of Medicine, Seoul, Korea.
出版信息
Korean J Parasitol. 1998 Sep;36(3):183-90. doi: 10.3347/kjp.1998.36.3.183.
Abstract
A cDNA expression library of Clonorchis sinensis adult worm was constructed, and screened out immunologically. One clone, pBCs31, was selected in view of its predominant reactivity with an experimentally infected rabbit serum. Recombinant C. sinensis antigen with 28 kDa as a beta-galactosidase fusion protein produced in Escherichia coli was identified by immunoblot analysis. The cloned gene was composed of 16 copies of a 30 base pair repeat and an additional 320 bases. The deduced amino acid sequence of the tandem repeat was AQPPKSGDGG. On RNA slot blot analysis. C. sinensis adult worm RNA showed a positive reaction with the cloned gene. Enzyme-linked immunosorbent assay using a purified recombinant antigen of pBCs31 showed high specificity for diagnosis of clonorchiasis.
摘要
构建了华支睾吸虫成虫的cDNA表达文库，并进行免疫筛选。鉴于一个克隆pBCs31与实验感染兔血清具有主要反应性，故将其挑选出来。通过免疫印迹分析鉴定了在大肠杆菌中产生的作为β-半乳糖苷酶融合蛋白的28 kDa重组华支睾吸虫抗原。克隆的基因由16个30碱基对重复序列和另外320个碱基组成。串联重复序列推导的氨基酸序列为AQPPKSGDGG。在RNA斑点印迹分析中，华支睾吸虫成虫RNA与克隆基因呈阳性反应。使用纯化的pBCs31重组抗原进行的酶联免疫吸附测定显示对华支睾吸虫病诊断具有高特异性。
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