酶法测定糖化血红蛋白的方法评价及标本前处理对结果的影响

Methodological evaluation on enzymatic method for detecting HbA1c and influence of sample pre-processing on detection result

目的探讨酶法测定糖化血红蛋白（HbA1c）方法学性能及其影响因素。方法采用酶法测定HbA1c,评价该方法学的精密度、抗干扰性、回收率、准确性以及标本前处理（抗凝、保存、离心）对结果的影响,分析与高效液相法（HPLC）相关性及偏倚程度。结果酶法批内高、中、低值变异系数（CV）为1.04%、1.26%、1.37%,批间为1.83%、2.24%、2.64%,与HPLC法呈线性相关（r=0.996,P〈0.01）;HbA1c靶值浓度为5.20%、6.40%、7.60%、8.80%、10.00%、11.20%,其回收率分别为100.15%、98.91%、98.84%、98.20%、103.62%、99.82%;当葡萄糖小于15.50 mmol/L、尿酸小于516.00μmol/L、胆红素小于217.00μmol/L、三酰甘油小于10.20mmol/L、尿素小于11.50mmol/L、清蛋白小于50g/L、球蛋白小于50g/L时,对结果无明显干扰。肝素钠、乙二胺四乙酸二钾（EDTA-K2）、枸橼酸钠抗凝标本HbA1c结果在-20-20℃保存3d无明显改变（P〉0.05）;标本500、1 000r/min（R=15cm）离心不同时间（1、2、5、10min）以及2 000r/min离心1min,其检测结果与3 000r/min离心5min比较,差异有统计学意义（P〈0.05）。结论酶法测定HbA1c其精密度、抗干扰性、准确性、线性范围均符合临床要求,与常规方法相比其相关性良好且偏差较小,可完全满足临床对HbA1c检测需求。

Objective To evaluate the methodological performance of the new enzymatic method for detecting glycated hemoglobin（HbAIc）and its influencing factors.Methods HbAIc was detected by the enzymatic method.The precision,anti-interference,recovery rate,accuracy and the influence of pre-processing（anti-coagulation,preservation,centrifugation）on the detection results were evaluated,its correlation with HPLC and the bias degree were analyzed.Results The within-run coefficients of variation（CVs）for high,middle and low value QC samples in the enzymatic assay were 1.04%,1.26% and 1.37%respectively and the between-run CVs were 1.83%,2.24% and 2.64%,respectively;the enzymatic method showed the linear correlation with HPLC（r=0.996,P〈0.01）;the HbA1 ctarget value concentrations were 5.20%,6.40%,7.60%,8.80%,10.00% and 11.20%respectively,the recovery rates were 100.15%,98.91%,98.84%,98.20%,103.62% and 99.82% respectively;the interference test showed that this method had no significant interference on the detection results when glucose15.50mmol/L,UA516.00μmol/L,bilirubin 217.00μmol/L,triglyceride10.20mmol/L,urea11.50mmol/L,albumin50g/L and globulin 50g/L.The HbA1 c detection results in the samples with anti-coagulation by heparin sodium,EDTA-2Kand sodium citrate stored for 3dunder-20-20 ℃ had no obvious change（P〉0.05）;the sample was centrifuged at 500,1 000r/min（R=15cm）for different time（1,2,5,10min）and at 2 000r/min for 1min,their detection results had statistical differences compared with the sample centrifuged=3 000r/min for 5min（P〈0.05）.Conclusion The precision,anti-interference,accuracy and linearity range of the enzymatic method all conform to the clinical requirement.Compared with the conventional method,its correlation is good with small deviation,which can entirely satisfy the demand of the HbAIc detection in clinic.