甲状旁腺激素荧光免疫层析定量检测技术的研制及应用研究  

The study and application on the parathyroid hormone fluorescence microsphere-immunochromatography quantification technique

作　　者：朱利国[1] 邹贤[1] 范俊[2] 周锦 黄飚[2] 

机构地区：[1]江苏省原子医学研究所附属江原医院,无锡214063 [2]江苏省原子医学研究所,无锡214063

出　　处：《现代免疫学》2018年第1期31-35,共5页Current Immunology

基　　金：无锡市科技支撑计划(CSE01N1239);江苏省卫生与计划生育委员会课题(Z201506)

摘　　要：研制一种操作简易、灵敏度高的荧光免疫层析试纸条,用于定量检测甲状旁腺激素(parathyroid hormone,PTH)。采用双抗体夹心法和荧光免疫层析技术,将标记抗体LA001与荧光微球结合于聚酯膜结合垫上,检测抗体CA001包被在硝酸纤维素膜上作为检测线(T),另将质控抗体(羊抗鼠IgG)包被在相互平行的硝酸纤维素膜上作为质控线(C),过量的鼠源标记的荧光微球在质控线处聚集产生质控荧光信号,待测样中的PTH在T处形成包被抗体-PTH抗原-标记抗体的夹心复合物并产生荧光信号,通过荧光检测仪计算出定量结果。试纸条检测PTH灵敏度最低可达0.8pg/mL,与促甲状腺激素(thyroid stimulating hormone,TSH)无交叉反应,不同批次试剂4 oC放置6个月稳定性好,批内变异系数(coefficient of variation,CV)为5.32%,批间CV值为6.74%。与电化学发光免疫法(electrochemiluminescence immunoassay,ECLIA)比较相关性较好,R2=0.993,差异无统计学意义,甲状旁腺组织结果与非甲状旁腺组织结果则有明显差异(P<0.01)。采用该荧光免疫层析试纸条,不仅可以提供较高的灵敏度和特异度,而且操作简单,适合临床工作的需要。To develop a simple and high sensitive fluorescence immunoassay strip for clinical use, we employed dual antibody sandwich method and fluorescence immune-chromatography for measuring the antigen. The fluorescence labeled antibody mi- crospheres LA001 were coated on polyester membrane-bound pad. Detection antibody CA001 was coated on the nitrocellulose membrane as the test line（T）, the quality control antibody （goat-anti-mouse IgG） was coated in nitrocellulose membrane in parallel as the control line（C）, excess microspheres coated with mouse antibody can gather in the control line（C）, which coated with anti-mouse antibody and the parathyroid hormone （PTH） in samples at the test line,could be the formation of antibody- antigen-antibody sandwich complexes and generate a fluorescent signal. Quantitative results could be automatically calculated by fluorescence detector. The results showed that the sensitivity of the test strip is 0.8 pg/mL and has no cross-reaction with TSH;the reagent has good stability for 6 months at 4℃. Intra-assay CV was 5.32% and inter-assay CV was 6.74%. Compared with the electrochemiluminescence immunoassay （ECHA） method, the correlation is good and the difference is not statistically significant. In conclusion, the fluorescent immune-chromatographic test strip provides high sensitivity, specificity,simple operation, low cost and is suitable for clinical detection.

关 键 词：甲状旁腺激素 甲状腺癌 荧光免疫层析 

分 类 号：R392.11[医药卫生—免疫学]