血浆中总同型半胱氨酸的测定：雅培IMx免疫测定法与高效液相色谱法的比较

Determination of total homocysteine in plasma: comparison of the Abbott IMx immunoassay with high performance liquid chromatography.

作者信息

Zighetti Maddalena Loredana, Chantarangkul Veena, Tripodi Armando, Mannucci Pier Mannuccio, Cattaneo Marco

机构信息

Department of Internal Medicine, Angelo Bianchi Bonomi Hemophilia and Thrombosis Center, University of Milan, via Pace 9, 20122 Milan, Italy.

出版信息

Haematologica. 2002 Jan;87(1):89-94.

PMID:11801469

Abstract

BACKGROUND AND OBJECTIVES

The aim of this study was to compare the performance of a commercially available IMx immunoassay with that of a reversed-phase high performance liquid chromatography (HPLC) method for measuring plasma total homocysteine (tHcy).

METHODS

The levels of tHcy before and after oral methionine loading (ML) were measured in 135 healthy subjects and 39 patients scheduled for routine tHcy determination. The IMx method uses fluorescence polarization immunoassay (FPIA) technology. The HPLC-method includes derivatization with ABD-F and post-column fluorescence detection.

RESULTS

The imprecision was very low with both methods for both normal (11 mmol/L) and high (29 mmol/L) tHcy levels. The within and between-run coefficients of variation were < 5%. Both methods were able to discriminate between similar concentrations of tHcy both at normal and moderately high levels. There was a good correlation between measurements obtained with the two methods (r = 0.985, p = 0.001). The mean levels of tHcy measured with the IMx assay tended to be slightly higher than those with the HPLC both in the fasting state (mean difference = 0.8 mmol/L) and after ML (5.3 mmol/L). However only the difference in post-ML levels was statistically significant (p < 0.001). The percentage of patients with hyperhomocysteinemia identified with the two methods was similar.

INTERPRETATION AND CONCLUSIONS

The IMx method compares well with an established HPLC method for measurement of fasting tHcy plasma levels.

摘要

背景与目的

本研究旨在比较市售IMx免疫分析法与反相高效液相色谱法（HPLC）测定血浆总同型半胱氨酸（tHcy）的性能。

方法

对135名健康受试者和39名计划进行常规tHcy测定的患者，测量口服蛋氨酸负荷（ML）前后的tHcy水平。IMx方法采用荧光偏振免疫分析（FPIA）技术。HPLC法包括用ABD-F衍生化和柱后荧光检测。

结果

对于正常（11 mmol/L）和高（29 mmol/L）tHcy水平，两种方法的不精密度都非常低。批内和批间变异系数均<5%。两种方法在正常和中度高水平下都能够区分相似浓度的tHcy。两种方法测得的结果之间有良好的相关性（r = 0.985，p = 0.001）。在禁食状态下，IMx分析法测得的tHcy平均水平往往略高于HPLC法（平均差异 = 0.8 mmol/L），ML后也是如此（5.3 mmol/L）。然而，只有ML后水平的差异具有统计学意义（p < 0.001）。两种方法识别出的高同型半胱氨酸血症患者百分比相似。