检测α-synuclein ELISA夹心法的建立  

Establishment of a sandwich ELISA for α-synuclein detection

作　　者：胡国艳[1] 罗新妮[2] 何柯新[1] 麦静雯[1] 侯乐[2] 钟笑梅[2] 宁玉萍[2] 刘学军[1] 

机构地区：[1]广州市精神病医院检验科,510370 [2]广州市精神病医院神经内科,510370

出　　处：《免疫学杂志》2013年第8期702-704,709,共4页Immunological Journal

基　　金：国家自然科学基金项目(30970902);广东省科技计划项目(2010B031600018);广州市科技计划项目(2010Y1-C631)

摘　　要：目的建立定量检测α-synuclein的ELISA夹心法。方法采用抗人α-synuclein单抗包被酶标板,以兔抗人α-synuclein多抗为夹心抗体、HRP标记羊抗兔IgG为检测抗体、重组人α-synuclein为标准品,建立检测α-synuclein的间接ELISA法,并对36例正常对照、27例路易体痴呆患者脑脊液样本进行了检测。结果建立的夹心ELISA法检测α-synuclein的线性范围为1.56～200 ng/ml,批内、批间变异系数分别为为7.51%和13.7%,36例正常对照、27例路易体痴呆患者脑脊液α-synuclein的含量比较差异有统计学意义(P<0.05)。结论成功建立了一种检测α-synuclein的夹心ELISA法。To establish an ELISA method for the detection of α-synuclein, the anti-human α-synuclein monoclonal antibodies and rabbit anti-human α-synuc｜ein poiyclonal antibodies were used as coating antibody and sandwich antibodies, respectively. Recombinated human α-synuclein as standard substance and HRP labeled goat anti-rabbit IgG as detector antibody, thus a sandwich ELISA quantitative immunological detection method for α- synuclein was established. Then the established method was used to determine α-synuclein level in 36 healthy volunteers and 27 Dementia of Lewy bodies （DLB） patients. The linear range of the detection was from 1.56 ng/ml to 200 ng/ml, while the coefficients of variation of within assay and between assays were 7.51% and 13.7%, respectively. This method showed that the concentration of α-synuclein in the CSF of DLB patients was lower than that in healthy human. All the result indicated that a stable sandwich ELISA for the detection of human α- synuclein was developed successfully, which would be helpful to further study the role of α-synuclein in neurodegenerative disease, and set a foundation for development a commercial assay kit.

关 键 词：ELISA 夹心法 Α-SYNUCLEIN 路易体痴呆 

分 类 号：R446.6[医药卫生—诊断学]