Determination of Human Granulocyte Elastase by the Immunoactivation Method on the Hitachi® 717 Automated Analyser
Determination of Human Granulocyte Elastase by the Immunoactivation Method on the Hitachi® 717 Automated Analyser
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在 Hitachi® 717 自动分析仪上通过免疫激活法测定人粒细胞弹性蛋白酶
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发表时间：
1991
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通讯作者：
W. Prellwitz
中科院分区：
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作者：
G. Hafner;M. Dreher;M. Lütgehaus;W. Ehrenthal;A. Heubner;H. Swars;W. Prellwitz
本文介绍了一种全机械化均相免疫分析法，采用免疫活化法快速、特异地测定血浆中的人粒细胞弹性蛋白酶（EC 3.4.21.37）。该方法使用来自绵羊的抗弹性蛋白酶抗体片段，与辣根过氧化物酶结合。这些酶-抗体缀合物与血浆中存在的弹性蛋白酶-α 1-蛋白酶抑制剂复合物结合。使用与辣根过氧化物酶偶联的非特异性绵羊抗体片段的单独样品空白校正样品基质引入的误差。使用临床化学分析仪Hitachi 717进行测量。单次测定可在10分钟内完成，需要24微升样品体积。测量范围约为20至1000微克/升弹性蛋白酶。对于运行内精密度，弹性蛋白酶浓度为45.7、89.1和385.4 μ g/L时的变异系数分别为4.77%、4.48%和1.85%;对于日间精密度，弹性蛋白酶浓度为31.1、65.5和440.2 μ g/L时的变异系数分别为15.81%、7.19%和4.12%。结果与非均相免疫测定结果的相关性（y = bx + a）显示出良好的一致性（r = 0.93，B = 1.11，a =-27.0，N = 121）。未观察到内源性物质和治疗剂量药物的干扰。使用215名健康个体的血浆（C-反应蛋白低于5 mg/l，白细胞计数4-8 x 10（9）/l）确定的参考区间为9-56 μ g/l（第2.5至第97.5百分位数），中位数为27 μ g/l。
This paper describes a fully mechanized homogeneous immunoassay using the immunoactivation method for the rapid and specific determination of human granulocyte elastase (EC 3.4.21.37) in plasma. The method uses anti-elastase antibody fragments from sheep, conjugated to horseradish peroxidase. These enzyme-antibody conjugates bind to the elastase-alpha 1-proteinase inhibitor complex present in plasma. A separate sample blank with non-specific sheep antibody fragments conjugated to horseradish peroxidase corrects for errors introduced by the sample matrix. Measurements were performed with the clinical chemistry analyser Hitachi 717. A single determination can be performed in 10 min, requiring 24 microliters sample volume. The measuring range is about 20 to 1000 micrograms/l elastase. For within-run precision the coefficients of variation are 4.77%, 4.48% and 1.85% for elastase concentrations of 45.7, 89.1 and 385.4 micrograms/l; for day-to-day precision the coefficients of variation are 15.81%, 7.19% and 4.12% for elastase concentrations of 31.1, 65.5 and 440.2 micrograms/l, respectively. Correlation (y = bx + a) of results with those from the heterogeneous immunoassay showed a good agreement (r = 0.93, b = 1.11, a = -27.0, N = 121). Interferences by endogeneous substances and by drugs at therapeutic doses were not observed. The reference interval, determined by using plasma from 215 healthy individuals (C-reactive protein less than 5 mg/l, leukocyte count 4-8 x 10(9)/l), was 9-56 micrograms/l (2.5th to 97.5th percentile), with a median of 27 micrograms/l.