糖化白蛋白酶法测定试剂研制与评价

目的：研发糖化白蛋白（GA）酶法测定试剂并进行方法学评价。方法：利用特异性的蛋白酶（PR）在3 min内裂解GA并释放出糖化氨基酸，然后利用酮胺氧化酶进行氧化并释放出过氧化氢，过氧化氢经Trinder’s反应显色后，与同样处理的标准溶液比较求得GA含量。结果：该试剂批内、批间不精密度（CV）分别为0.84%～1.88%和1.14%～2.35%；线性范围为1.3～1 200 μmol/L，血清样本2～16倍稀释范围与原倍血清的百分偏差在99.3%～108.6%之间，平均回收率为101.8%；血清中胆红素＜35 mg/dL，甘油三酯（TG）＜750 mg/dL，血红蛋白＜200 mg/dL，胆汁酸＜8 mg/dL，尿酸＜33 mg/dL，葡萄糖＜1 800 mg·dL-1，维生素C 15 mg/dL，EDTA·K2 2.5 mg/dL，肝素钠80 U/L，枸橼酸钠8.0 mg/dL，对550 μmol/L GA的检测干扰＜±5%。与日本ASAHI公司提供的GA测定试剂方法进行比较，回归方程及相关系数分别为Y=0.9991X+ 0.0809，r=0.9958；试剂置4 ℃开盖储存至少稳定30 d，置4 ℃加盖储存至少稳定12个月。结论：GA酶法测定试剂精密度良好，线性范围宽，稀释准确性满足临床需要，回收率较高，抗干扰能力较强，与进口试剂相关性好，具有推广应用价值。

The development and evaluation of glycated albumin enzymatic determination reagent

Objective: To develop and evaluate the serum glycated albumin enzymatic determination reagent. Methods: The analysis was based on the fact that the glycated albumin glycated albumin was hydrolyzed to glycated amino acids by special proteinase digestion, and glycated amino acids were oxidized to produce hydrogen peroxide, which was quantitatively measured through the Trinder’s chromogenic reaction, compared with the same treatment of the standard solution. Results: The within-run and between-run CVs of this reagent were 0.84%～1.88% and 1.14%～2.35%, respectively; the method was liner in the range of 1.3～1 200 μmol•L-1, percent deviation of the measured results of diluted serum samples diluted by 2 to 16 times compared to the results of the original samples ranged from 99.3%～108.6%, the average rate of recovery was 101.8%; No significant interference (bias%≤±5%) was found in the test of 550 μmol•L-1 GA analysis when bilirubin <35 mg•dL-1, triglyceride<750 mg•dL-1, hemoglobin <200 mg•dL-1, bile acid <8 mg•dL-1, uric acid <33 mg•dL-1, glucose <1800 mg•dL-1, Vitamin C <15 mg•dL-1, EDTA•K2 <2.5 mg•mL-1, Heparin Sodium <80 U•L-1, Sodium citrate <8.0 mg•mL-1. Comparing this methods (Y) with the ASAHI method (X), the regression equations is Y=0.9991X+0.0809, r=0.9958. The reagent was stable at least 30 days when it was open cover stored and at least 12 months when it was sealed stored at 4 ℃. Conclusion: This reagent shows high precision, wide linear range, and the dilution accuracy can meet the clinical needs. It also shows high recovery, good anti-interference capability, good correlation with imported reagent, and suitable for clinical application.