新型量子点荧光免疫层析法测定血清淀粉样蛋白A的应用研究  

Application study of novel quantum dot fluorescence immunochromatography for the determination of serum amyloid A

作　　者：张仙娜 玄光善[1] ZHANG Xianna;XUAN Guangshan(College of Chemical Engineering,Qingdao University of Science and Technology,Qingdao,Shandong 266042,China)

机构地区：[1]青岛科技大学化工学院,山东青岛266042

出　　处：《国际检验医学杂志》2024年第16期2010-2016,2022,共8页International Journal of Laboratory Medicine

摘　　要：目的探讨新型量子点荧光免疫层析法测定血清淀粉样蛋白A(SAA)的应用。方法采用双抗夹心法结合量子点荧光免疫层析技术制备以二硝基苯酚(DNP)-牛血清白蛋白(BSA)系统为质控线的SAA检测试剂盒。通过鸡IgY-羊抗鸡IgY系统作为对照评价了DNP-BSA系统作为质控线的可行性,并通过优化量子点微球与抗体的偶联条件,进一步提高试剂盒的性能,并对试剂盒的空白限、检出限、线性范围、精密度、准确度、特异度、稳定性,以及临床实际样本测定等方面进行了评价。结果以DNP-BSA系统作为质控线的SAA试剂盒受干扰成分浓度的影响小,稳定性高,热稳定性好。量子点微球与SAA抗体偶联比例为100μg/mg,偶联反应液浓度为25 mmol/L 3-吗啉丙磺酸pH 7.4,与DNP-BSA偶联比例为100μg/mg,偶联反应液浓度为25 mmol/L吗啉乙磺酸pH 6.0时偶联效果最佳。试剂盒最低检出限为0.5 mg/L;线性范围为1~200 mg/L,R2≥0.99;批内精密度变异系数(CV%)≤5.31%,批间精密度CV%≤15%;在低、高浓度的回收率分别为101.42%、98.83%;在血红蛋白浓度≤5 g/L,胆红素浓度≤0.15 g/L,胆固醇浓度≤15 g/L,甘油三酯浓度10 g/L时,SAA的检测结果无干扰;试剂盒在50℃放置21 d,稳定性良好;试剂盒与Roche的SAA电化学发光试剂盒同步检测67例样本结果具有高度一致性。结论研制的SAA检测试剂盒灵敏度、线性、精密度、准确度、特异度及加速稳定性都满足试剂盒的要求,与鸡IgY-羊抗鸡IgY系统相比DNP-BSA系统作为质控线独立性好,测定临床样本准确度和热稳定性更佳。Objective To investigate the application of novel quantum dot fluorescence immunochromatography for the determination of serum amyloid A(SAA).Methods Using the principle of double antibody sandwich method and quantum dots fluorescence immunochromatography technology to prepare SAA detection kit with Dinitrophenol(DNP)-Bovine serum albumin(BSA)system as the control line.The feasibility of using the DNP-BSA system as a quality control line was evaluated using the chicken IgY-sheep anti-chicken IgY system as a control,and further improve the performance of the reagent kit by optimizing the coupling conditions between quantum dot microspheres and antibodies.Evaluated the blank limit,detection limit,linear range precision,accuracy,specificity,stability,as well as clinical samples determination of the kit.Results The SAA kit using DNP-BSA system as the quality control line was less affected by the concentration of interfering components,had high stability,and good thermal stability.The optimal coupling effect was achieved when the coupling ratio between quantum dots and SAA antibodies was 100μg/mg,and the coupling reaction solution was 25 mmol/L 3-morpholine propyl sulfonate pH 7.4,and the coupling ratio with DNP-BSA was 100μg/mg and the coupling reaction solution was 25 mmol/L morpholine ethanesulfonate pH 6.0.The detection limit of SAA kit was 0.5 mg/L;the linear range was 1-200 mg/L,R^(2)≥0.99;the intra assay precision variation coefficient(CV%)≤5.31%,and the inter assay precision CV%≤15%;the recoveries in the low,and high concentrations were 101.42%and 98.83%,respectively.When hemoglobin concentration≤5 g/L,bilirubin concentration≤0.15 g/L,cholesterol concentration≤15 g/L,triglyceride concentration≤10 g/L,SAA detection results had no interference.The stability of the kit was good when it was stored at 50℃for 21 days.The kit is highly consistent with that of Roche's SAA electrochemiluminescence kit in the simultaneous detection of 67 samples.Conclusion The sensitivity,linearity,precision,accuracy,s

关 键 词：二硝基苯酚 牛血清白蛋白 量子点 荧光免疫层析 血清淀粉样蛋白A 

分 类 号：R446.6[医药卫生—诊断学]