均相酶免疫法检测血清维生素D含量的性能验证

Determination of 25-Hydroxyvitamin D by Homogeneous Enzyme Method

目的建立均相酶免疫法测定末梢血25-羟维生素D(25(OH)D)的方法。方法采用苏州博源医疗生产的均相酶法总25-羟维生素D检测试剂,验证其正确度、精密度、线性范围、可报告范围、灵敏度、方法比对及干扰试验等分析性能指标。结果四个不同浓度(8.00 ng/mL,17.50 ng/mL,35.00 ng/mL,70.00 ng/mL)25(OH)D校准品的偏倚分别为0.44%、-1.91%、1.51%和-0.23%;低值、高值血清样品的批内精密度和总精密度分别为2.22%、1.69%和2.42%、2.52%,小于厂商声明的1/4 TEa(6.25%)和1/3 TEa(8.33%);线性范围为8.0~70 ng/mL,可报告范围上限为210 ng/mL;分析灵敏度符合厂商声明标准(2.9 ng/mL)。方法学比对收集110例临床血清样品,分别使用均相酶免疫法(试验方法,Y)与罗氏发光(参比方法,X)的25(OH)D试剂进行检测,Deming回归方程为Y=0.9988X-0.8575,R2=0.9281。结论均相酶免疫法检测总25-羟维生素D试剂各项分析验证均符合要求,性能良好,满足临床应用要求。

Objective To establish a homogeneous enzyme method for measuring 25-hydroxyvitamin D(25(OH)D).Methods Total 25(OH)D detection reagent produced by Suzhou Boyuan was used to verify the accuracy,precision,linear range,reporting range,sensitivity and method comparison.Results The bias of four different concentrations(8.00 ng/mL,17.50 ng/mL,35.00 ng/mL,70.00 ng/mL)of 25(OH)D calibrators was 0.44%,-1.91%,1.51%,and-0.23%,respectively.The intra-batch precision and total precision of low-and high-value serum samples were 2.22%,1.69%and 2.42%,2.52%,respectively,less than the 1/4 TEa(6.25%)and 1/3 TEa(8.33%)stated by the manufacturer.The linear range was 8.0-70 ng/mL,the reportable range was 210 ng/mL,and the analytical sensitivity meets the vendor claim criteria(2.9 ng/mL).In comparison,110 clinical serum samples were collected and tested with 25(OH)D reagent by homogeneous enzyme immunoassay(test method,Y)and Roche luminescence(reference method,X).Deming regression equation was Y=0.9988X-0.8575,R2=0.9281.Conclusion The analysis of total 25-hydroxyvitamin D reagent meets the requirements and shows good performance,which can initially meet the requirements of clinical application.