荧光和比色信号检测骨髓过氧化物酶的四种免疫测定方法的比较。

I Janda, H Jaensch, J Braun, W G Wood
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引用次数: 6

摘要

本文介绍了体液中髓过氧化物酶测定的四种不同的测定系统。一种是基于传统的化学发光,两种是基于发光扩增酶的测量,使用蜘蛛烷-1,2-二氧乙烷与碱性磷酸酶或鲁米诺/过氧化物酶/4-碘酚偶联过氧化物酶标签。测定范围为0-600微克/升过氧化物酶。健康志愿者血浆中EDTA的参考范围上限为250微克/升(95%置信限)。如复合精度曲线所示,所有测定的测定内变异系数均小于5%。在整个浓度范围内,测定间差异小于10%。光产生的动力学曲线在2小时内进行增强发光测定。将这些测定的动态范围与使用4-硝基苯磷酸碱性磷酸酶的常规比色法进行比较。该试验使用市售的髓过氧化物酶制剂进行标准化,该制剂具有明确的酶活性。以质量为单位对蛋白质含量进行估计,并对实验室标准进行比较和校准。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A comparison of four immunometric assays for myeloperoxidase using luminescent and colorimetric signal detection.

This communication presents four different assay systems for the determination of myeloperoxidase in body fluids. One is based on conventional chemiluminescence, two on luminescence-amplified enzyme measurement using either spiroadamantane-1,2-dioxetanes with alkaline phosphatase or luminol/peroxidase/4-iodophenol coupled with a peroxidase label. The assays covered the range 0-600 micrograms/l peroxidase. Established reference range for EDTA plasma from healthy volunteers gave an upper limit of 250 micrograms/l (95% confidence limits). Intra-assay coefficients of variation were less than 5% for all assays, as seen in compound precision profiles. Inter-assay variation was less than 10% throughout the whole concentration range. Kinetic curves for the light production were performed over a 2 hour period for the enhanced luminescence assays. Dynamic ranges of these assays were compared with the conventional colorimetric assay using 4-nitrophenyl phosphate--alkaline phosphatase. The assay was standardized using a commercially available myeloperoxidase preparation with defined enzymatic activity. The protein content was estimated and the laboratory standard compared and calibrated in mass units.

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