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目的建立有关急性冠脉综合征(ACS)患者血清中妊娠相关血浆蛋白A(PAPP-A)的均相化学发光免疫检测方法。方法使用PAPP-A抗体与受体微球和生物素连接,以双抗体夹心法,生物素化抗体和受体微球与血清中PAPP-A结合,以鲁米诺氧途径免疫分析(LOCI)检测PAPP-A浓度。评价此方法各项性能指标。数据分析使用ELISA Calc软件。结果本研究PAPP-A检测方法灵敏度为0.55 m U/L,回收率为95%~100%;线性、稳定性及特异性良好;重复性批内变异系数(CV)<10%,批间CV<15%;检测不易受血红蛋白、甘油三酯和胆红素的干扰。该方法与时间分辨荧光免疫方法(TRFIA)进行相关分析(r=0.990,P<0.05)。检测104例正常健康人(男/女为43/61,年龄为24岁~69岁)血清PAPP-A,初步推算正常参考范围为1.60 m U/L~25.12 m U/L。结论本研究所建立的超敏PAPP-A检测方法准确性好,灵敏度、回收率、特异性较高,与TRFIA相关性良好,评价了其应用于临床检测的可行性,并为参考范围的确定提供了一定的依据。
Objective To establish a homogeneous chemiluminescence immunoassay for the detection of pregnancy-associated plasma protein A (PAPP-A) in the serum of patients with acute coronary syndrome (ACS). Methods PAPP-A antibody was conjugated to acceptor microspheres and biotin, and bound to PAPP-A in serum by double-antibody sandwich method, biotinylated antibody and acceptor microspheres. The luminol pathway immunoassay (LOCI) PAPP-A concentration was measured. Evaluation of the performance of this method. Data analysis using ELISA Calc software. Results The sensitivity of PAPP-A assay was 0.55 mU / L and the recovery was 95% -100%. The linearity, stability and specificity were good. The coefficient of variation (CV) was less than 10% <15%; detection is not susceptible to the interference of hemoglobin, triglycerides and bilirubin. The method was correlated with the time-resolved fluorescence immunoassay (TRFIA) (r = 0.990, P <0.05). The serum PAPP-A level of 104 healthy controls (male / female 43/61, age range from 24 to 69 years old) was detected. The normal reference range was 1.60 mU / L ~ 25.12 mU / L. Conclusion The established method for detecting the hypersensitive PAPP-A in this study has good accuracy, high sensitivity, good recovery, high specificity and good correlation with TRFIA. It evaluates the feasibility of its application in clinical detection and defines the reference range Provide a certain basis.