两种校准品和3种双缩脲试剂测定血清总蛋白结果偏倚评估及方法学评价

Bias Analysis and Methodological Evaluation on Test Results of Serum Total Protein with 2 Calibrators and 3 Biuret Reagents

目的评估两种校准品及3种双缩脲试剂盒测定血清总蛋白结果的偏倚,评价3种双缩脲试剂盒血清总蛋白的精密度、线性和灵敏度。方法用3种双缩脲试剂盒及两种校准品组合成6个检测系统,按NCCLS EP9-A文件,评估3种双缩脲试剂盒及两种校准品测定结果偏倚和统计学分析;按NCCLS EP5-A文件,评价3种双缩脲试剂盒批内、批间、日间和总CV%。按NCCLS EP6-P文件评价3种双缩脲试剂盒的线性;比较3种双缩脲试剂盒测定同一份血清在540nm处的吸光度。结果用Roche校准血清(c.f.a.s)校准仪器,双缩脲试剂盒B、C与试剂盒A比较,差异无统计学意义(P>0.05);用总蛋白标准液校准仪器,试剂盒B与试剂盒A比较,差异无统计学意义(P>0.05),试剂盒C与试剂盒A比较,差异具有统计学意义(P<0.01);3种双缩脲试剂盒测定血清总蛋白总CV为0.86%～1.16%;波长为540nm,试剂盒A的吸光度明显高于试剂盒B、C;试剂盒A和B线性良好(F<F0.05),试剂盒C不可接受(F>F0.05)。结论测定血清总蛋白,建议用Doumas配方的双缩脲试剂和基质状态与病人血清相似的校准血清。

Objective To analyze test bias of serum total protein with 2 calibrators and 3 biuret reagents, and to evaluate precision, linearity and sensitivity of the 3 biuret reagents and 2 calibrators . Methods Three biuret reagents and 2 calibrators consist of 6 test systems. According to EP9 -A （NCCLS） , test bias of 3 biuret reagents and 2 calibrators was analyzed statistically; according to EP5 -A, the results of intra - batch, between batch and batches on different days, and total CV% of 3 biuret reagents analyzed; according to EP6 -P, lincarity of 3 biuret reagents analyzed, 3 biuret reagents absorbance of the same serum at 540nm compared. Results The instrument was adjusted with Roche calibration serum （c. f. a. s）. Biuret reagent B compared with biuret reagent A, the difference was not significant statistically （ P 〉 0.05 ） ; reagent C compared with reagent A, the difference was significant statistically （ P 〈 0.01 ）. Total CV of serum total protein with 3 biuret reagents was 0.86% - 1.16% , wavelength 540nm, absorbance of reagent A was significantly higher than reagent B and reagent C. Reagent A and reagent B was of good linearity （F 〈 F0.05）. Reagent C couldn＇t be accepted （F 〉 F0.05 ）. Conclusion We recommend biuret reagents of Doumas compounding and its base state, and calibration serum similiar to patients serum for test of serum total protein.