【摘要】 目的 建立一种定量检测血清中β淀粉样蛋白1-42(Aβ1-42)的磁微粒化学发光免疫检测方法.方法 制备活化磁性微球，并标记抗Aβ1-42抗体；制备Aβ1-42抗体-吖啶酯发光试剂，建立一步式定量检测Aβ1-42浓度的磁微粒化学发光免疫检测方法，组装成试剂盒，并评估其灵敏度、准确度、重复性、特异性、稳定性等，推测血清Aβ1-42正常参考区间.结果 该方法对血清Aβ1-42检测灵敏度评估的空白限(LoB)为0.03 pg/mL,检测限(LoD)为0.05 pg/mL,功能灵敏度(FS)为0.11 pg/mL,线性范围0.1～5 000 pg/mL,加标回收率为103.78%～106.10%;试剂盒检测的分析内精密度为1.06%～2.70%,分析间精密度为3.61%～5.21%,总不精密度为5.93%～8.60%.在试剂盒特异性方面，与血清中常见蛋白的交叉反应率低于2%;同时，样本中常见的3种干扰物在胆红素、甘油三酯、血红蛋白浓度分别低于40 U/L、2 mmol/L、3 g/L时，检测结果基本不受影响.试剂盒在37℃下可稳定保存7 d以上，说明2～8℃下产品有效期不低于12个月.且100例正常人血清检测所得Aβ1-42正常参考区间>654.2 pg/mL.结论 本方法建立的Aβ1-42磁微粒化学发光免疫检测方法具有灵敏度高、准确性强、重复性和特异性高等优点，可为血清中Aβ1-42精准定量检测提供新的技术手段.更多还原

【Abstract】 Objective To establish a magnetic particle chemiluminescence immunoassay detection method for quantitative detection of amyloid beta 1-42(Aβ1-42) in serum.Method Activated magnetic microspheres and labeled anti-Aβ1-42 antibodies, meanwhile prepared Aβ1-42 paird antibodies-acridinium ester conjugates, and then established a one-step magnetic bead chemiluminescence detection method for quantitatively detecting the concentration of Aβ1-42 levels.The kit was assembled and evaluated for sensitivity, accuracy, reproducibility, specificity, stability, etc.The normal reference interval of Aβ1-42 was inferred.Results The blank limit(LoB) of the method was 0.03 pg/mL for the sensitivity detection of Aβ1-42 in serum.LoD was 0.05 pg/mL,the sensi-tivity of this kit was 0.11 pg/mL,and the linear range was 0.1～5 000 pg/mL;The recovery rate of the intra-assay was between 103.78% and 106.10%;The intra-assay and inter-assay of kits were 1.06%～2.70%,3.61%～5.21% respectively.The cross-reaction rates of common proteins in serum were all less than 2%;At the same time, when the concentration of bilirubin, triglyceride and hemoglobin were not higher than 40 U/L,2 mmol/L and 3 g/L,the results of detection were not affected.It can be stored stably at 37 ℃ for more than 7 days, the term of validity of 2～8 ℃ product is not less than 12 months.The normal reference interval of Aβ1-42 in 100 normal human serum was more than 654.2 pg/mL.Conclusion This study established Aβ1-42 magnetic particle chemiluminescence immunoassay, which has the advantages of high sensitivity, high accuracy, high repeatability and specificity, it can provide a accurate quantitative detection method for Aβ1-42 levels in serum.更多还原