快速定量检测血清α-羟丁酸脱氢酶干化学方法的建立

摘要

Objective:To establish a rapid and quantitative dry chemistry method to detectα-hydroxybutyrate dehydrogenase(α-HBDH) in serum.Methods:Test strip was designed,the structure of test strip consisted of a spreading layer,a reagent layer and a supporting layer.Test strips and DC-101dry biochemistry analyzer constitute a detection system.The precision,accuracy,linear range,stability of the test strips and methodological comparison were evaluated according to the clinical test method evaluation schemes EP5-A2,EP15-A,and EP6-A provided by CLSI.Results:The intra coefficient of variation of test strips for Randox low and high level quality controls were 4.1％ and 5.8％ respectively,which were lower than the standard of 1/4CLIA'88.The relative bias of test were 4.9％ and 5.3％ respectively using 267U/ L and 465U/L Randox calibrators within 1/2TEa.The linear range was 20～1000U/L(r2=0.992).Test strips were valid for 12months.The slope of Passing-Bablock regression equation was 0.995which was close to the theoretical value 1.Bland-Altman analysis showed that 4.16％ of the points were outside the consistency limit.Conclusion:Prepared test strips could be well applied to detectα-HBDH,which could meet the needs of clinical diagnosis.%目的:建立一种快速定量检测人体血清α-羟丁酸脱氢酶(α-HBDH)的干化学检测方法.方法:制备试纸,试纸由扩散层、试剂层、支持层组成,试纸和DC-101干式生化分析仪组成检测系统,参照CLSI提供的临床检验方法评价方案EP5-A2、 EP15-A、EP6-A,对试纸的精密度、准确度、线性范围、稳定性、方法学比对等性能进行评价.结果:试纸测定朗道低、高值质控品的批内变异系数分别为4.1％和4.6％,小于1/4CLIA''88指标;对267U/L、465U/L两个浓度朗道校准品测定相对偏差分别为4.9％、5.3％,在1/2TEa范围内;线性范围在20～1!000U/L(r2=0.992);试纸有效期12个月;Passing-Bablok回归方程的斜率为0.995,接近理论值1;Bland-Altman分析显示4.16％的点在一致性界限之外.结论:所制备的试纸可应用于α-HBDH的检测,能够满足临床需求.