权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

The establishment of serum diagnostic method for canine myasthenia gravis

犬重症肌无力血清诊断方法的建立

基本信息

批准号：
15580285
负责人：
UZUKA Yuji
金额：
$ 2.05万
依托单位：
Gifu University (2004)Obihiro University of Agriculture and Veterinary Medicine (2003)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15580285/
关键词：
myasthenia gravis acetylcholine receptor canine insect cell expression α-subunit fetal type of AChR mature type of AChR ELISA αサブユニット εサブユニットコスミックII TE671 無症筋無力症 α-サブユニット

项目摘要

Acquired myasthenia gravis (MG) is the autoimmune neuromuscular disease due to the anti-acetylcholine receptor (AChR) antibody for the acetylcholine receptor (AChR) on postsynaptic menbrane of skeletal muscle. Anti-AChR antibodies are cllasfied by blocking antibody recognized acetylcholine binding site and binding antibody recognized main immunogenic region. In this study, the gene encoding the entire canine AChR α-subunit was cloned and inserted into an Escherichia coli expression vector and an insect cells expression vector, and subsequently expressed recombinant canine AChR α subunit pretein (re-AchR α P) respectively. Western blot analyses and ELISA for serological diagnosis of acquired MG were carried out using the re-AChR α p expressed in E. coli and in insect cells severally. The re-AChR α p was able to confirm severally, but these immunoassay with these proteins was not able to differentiate between sera from antibody-positive MG dogs and sera from antibody-negative healthy dogs. Therefore, at present stage, this assay is not useful for the clinical diagnosis of acquired MG.In additon, we investigated the evaluation of the measuring device of anti-AChR antibody for human sera (Cosmic II), which was utilized AChR a P from rhabdomyosarcoma as a antigen. 4 measured values were relatively low and 6 were high compared from conventional measuring method in 14 canine sera, which were procured from clinical acquired MG. This implies the decreased sensitivity by different species specificity and the reinforcement sensitivity by large quantities of mature type AChR of

获得性重症肌无力（MG）是由于骨骼肌突触后膜上乙酰胆碱受体（AChR）的抗乙酰胆碱受体（AChR）抗体引起的自身免疫性神经肌肉疾病。抗achr抗体分为阻断抗体识别的乙酰胆碱结合位点和结合抗体识别的主要免疫原区。本研究克隆了整个犬AChR α-亚基的编码基因，分别插入大肠杆菌表达载体和昆虫细胞表达载体，分别表达重组犬AChR α亚基蛋白（re-AchR α P）。利用大肠杆菌和昆虫细胞中分别表达的re-AChR α p进行免疫印迹（Western blot）和ELISA检测获得性MG的血清学诊断。re-AChR α p能够分别得到证实，但用这些蛋白进行的免疫测定不能区分抗体阳性MG犬的血清和抗体阴性的健康犬的血清。因此，在目前阶段，这种检测方法对获得性MG的临床诊断还不是很有用。此外，我们还研究了以横纹肌肉瘤AChR a P为抗原的人血清抗AChR抗体测定装置（Cosmic II）的评价。14份临床获得性MG犬血清中，与常规测定方法相比，4份测定值较低，6份测定值较高。这说明不同的物种特异性会降低敏感性，而大量成熟型AChR会增强敏感性