人血清淀粉样蛋白A磁微粒化学发光免疫分析定量检测方法的建立及性能评价

Establishment and Performance Evaluation of the Quantitative Detection for Human Serum Amyloid A Based on Chemiluminescent Immunoassay

目的建立人血清淀粉样蛋白A(human serum amyloidA,SAA)磁微粒化学发光免疫分析定量检测方法。方法采用基于磁微粒化学发光免疫分析的双抗体夹心法定量检测人血液中SAA含量。评价该方法学的空白限、精密度、线性、回收率和钩状效应等分析性能指标,并用以上方法检测60例细菌或病毒感染患者血清和40例正常人血清,进行临床比对试验研究。结果该方法的空白限为0.1mg/L;分析内精密度不高于5%,分析间精密度不高于10%;在2~200 mg/L范围内,线性相关系数可达0.990;准确度偏倚不高于15%;回收率在100%±15%以内;样本中SAA的含量为10000mg/L时,未发现钩状效应;与西门子免疫比浊法试剂平行比对试验,回归方程为:Y=1.0998X-2.6739,相关系数r2为0.9834。结论该方法灵敏度高、精密度好、线性范围宽和临床相关性好,具有一定的临床应用价值。

Objective To establish a quantitative detection method for human serum amyloid A(SAA)by chemiluminescence immunoassay.Methods The double antibody sandwich method based on magnetic particle chemiluminescence immunoassay was used to quantitatively detect the SAA content in human blood.The analytical performance of the established assay was further evaluated by a series of analysis,such as blank limit,precision,linearity,recovery rate and hook effect.The serum of 60 bacterial or viral infected patients and 40 normal human serum were detected by this method for clinical comparison test.Results The limit of blank was 0.1 mg/L.The within variance was lower than 5 percents,and days varican was lower than 10 percents.The linear range was 2~200 mg/L,and the correlation coefficient(r)was 0.990.Measurement relative bias for trueness was less than 15%.The spiking recovery for accuracy was(100±15)%,and there was no hook effect when the sample with 10 000 mg/L SAA.The determination coefficient r^2 was 0.983 4 in method comparison,regression equation was Y=1.099 8X-2.673 9.Conclusion The established human SAA magnetic particle chemiluminescence immunoassay quantitative detection method has high sensitivity,fast detection speed,specificity,high clinical diagnostic value.