Sako Y, Nakao M, Ikejima T, Piao X Z, Nakaya K, Ito A
Department of Parasitology, Asahikawa Medical College, Asahikawa, Japan.
J Clin Microbiol. 2000 Dec;38(12):4439-44. doi: 10.1128/JCM.38.12.4439-4444.2000.
Neurocysticercosis (NCC) caused by infection with the larvae of Taenia solium is an important cause of neurological disease worldwide. In order to establish an enzyme-linked immunosorbent assay (ELISA) for this infection using recombinant proteins, we carried out molecular cloning and identified four candidates as diagnostic antigens (designated Ag1, Ag1V1, Ag2, and Ag2V1). Except for Ag2V1, these clones could encode a 7-kDa polypeptide, and Ag2V1 could encode a 10-kDa polypeptide. All of the clones were very similar. Except for Ag2V1, recombinant proteins were successfully expressed using an Escherichia coli expression system. Immunoblot analysis of NCC patient sera detected recombinant proteins, but because reactivity to recombinant Ag1 was too weak, Ag1 was not suitable as an immunodiagnostic antigen. So, Ag1V1 and Ag2 were chosen as ELISA antigens, and the Ag1V1/Ag2 chimeric protein was expressed. Of 49 serum samples from NCC patients confirmed to be seropositive by immunoblot analysis, 44 (89.7%) were positive by ELISA. No assays of serum samples from patients with other parasitic infections recognized the Ag1V1/Ag2 chimeric protein. The Ag1V1/Ag2 chimeric protein obtained in this study had a high value for differential immunodiagnosis.
由猪带绦虫幼虫感染引起的神经囊尾蚴病（NCC）是全球神经系统疾病的一个重要病因。为了利用重组蛋白建立针对这种感染的酶联免疫吸附测定（ELISA），我们进行了分子克隆，并鉴定出四种候选诊断抗原（命名为Ag1、Ag1V1、Ag2和Ag2V1）。除Ag2V1外，这些克隆可编码一种7 kDa的多肽，而Ag2V1可编码一种10 kDa的多肽。所有克隆都非常相似。除Ag2V1外，利用大肠杆菌表达系统成功表达了重组蛋白。对NCC患者血清的免疫印迹分析检测到了重组蛋白，但由于对重组Ag1的反应性过弱，Ag1不适合作为免疫诊断抗原。因此，选择Ag1V1和Ag2作为ELISA抗原，并表达了Ag1V1/Ag2嵌合蛋白。在通过免疫印迹分析确认血清学阳性的49份NCC患者血清样本中，44份（89.7%）通过ELISA检测为阳性。其他寄生虫感染患者的血清样本检测均未识别出Ag1V1/Ag2嵌合蛋白。本研究获得的Ag1V1/Ag2嵌合蛋白在鉴别免疫诊断方面具有很高的价值。
