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使用衰减全反射傅里叶变换红外光谱法检测人类指甲剪屑中的糖基化,一种用于糖尿病诊断和监测的新标志物。

Glycation in human fingernail clippings using ATR-FTIR spectrometry, a new marker for the diagnosis and monitoring of diabetes mellitus.

作者信息

Coopman Renaat, Van de Vyver Thijs, Kishabongo Antoine Sadiki, Katchunga Philippe, Van Aken Elisabeth H, Cikomola Justin, Monteyne Tinne, Speeckaert Marijn M, Delanghe Joris R

机构信息

Department of Clinical Chemistry, Ghent University Hospital, Gent, Belgium.

Department of Laboratory Medicine, Catholic University of Bukavu, Bukavu, Democratic Republic of the Congo.

出版信息

Clin Biochem. 2017 Jan;50(1-2):62-67. doi: 10.1016/j.clinbiochem.2016.09.001. Epub 2016 Sep 4.

Abstract

OBJECTIVES

Although HbA1c is a good diagnostic tool for diabetes, the precarity of the health system and the costs limit the use of this biomarker in developing countries. Fingernail clippings contain ±85% of keratins, which are prone to glycation. Nail keratin glycation may reflect the average glycemia over the last months. We explored if attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) can be used as a non-invasive tool for assessing glycation in diabetes.

DESIGN AND METHODS

Using ATR-FTIR spectroscopy, glycation and deglycation experiments with fructosamine 3-kinase allowed to identify the spectrum that corresponds with keratin glycation in fingernail clippings. Clippings of 105 healthy subjects and 127 diabetics were subjected to the standardized ATR-FTIR spectroscopy method.

RESULTS

In vitro glycation resulted in an increased absorption at 1047cm. Following enzymatic deglycation, this peak diminished significantly, proving that the AUC between 970 and 1140cm corresponded with glycated proteins. Within-run CV of the assay was 3%. Storage of nail clippings at 37°C for 2weeks did not significantly change results. In diabetics, glycated nail protein concentrations (median: 1.51μmol/g protein, IQR: 1.37-1.85μmol/g protein) were significantly higher than in the controls (median: 1.19μmol/g protein, IQR: 1.09-1.26μmol/g protein) (p<0.0001). ROC analysis yielded an AUC of 0.92 at a cut-off point of 1.28μmol/g nail (specificity: 82%; sensitivity: 90%). No correlation was observed between the glycated nail protein concentrations and HbA1c.

CONCLUSIONS

Protein glycation analysis in fingernails with ATR-FTIR spectroscopy could be an alternative affordable technique for diagnosing and monitoring diabetes. As the test does not consume reagents, and the preanalytical phase is extremely robust, the test could be particularly useful in developing countries.

摘要

目的

尽管糖化血红蛋白(HbA1c)是诊断糖尿病的良好工具,但卫生系统的不稳定以及成本限制了该生物标志物在发展中国家的使用。指甲剪屑中约含85%的角蛋白,角蛋白易于发生糖基化。指甲角蛋白糖基化可能反映过去几个月的平均血糖水平。我们探讨了衰减全反射傅里叶变换红外光谱(ATR-FTIR)是否可作为评估糖尿病患者糖基化的非侵入性工具。

设计与方法

使用ATR-FTIR光谱,通过果糖胺3-激酶进行糖基化和去糖基化实验,以确定与指甲剪屑中角蛋白糖基化相对应的光谱。对105名健康受试者和127名糖尿病患者的指甲剪屑采用标准化的ATR-FTIR光谱法进行检测。

结果

体外糖基化导致在1047cm处吸收增加。酶促去糖基化后,该峰显著降低,证明970至1140cm之间的曲线下面积(AUC)与糖基化蛋白相对应。该检测方法的批内变异系数(CV)为3%。将指甲剪屑在37°C下保存2周并未显著改变检测结果。糖尿病患者指甲糖基化蛋白浓度(中位数:1.51μmol/g蛋白,四分位间距:1.37 - 1.85μmol/g蛋白)显著高于对照组(中位数:1.19μmol/g蛋白,四分位间距:1.09 - 1.26μmol/g蛋白)(p<0.0001)。受试者工作特征(ROC)分析在指甲糖基化蛋白浓度为1.28μmol/g的截断点处得出AUC为0.92(特异性:82%;敏感性:90%)。未观察到指甲糖基化蛋白浓度与HbA1c之间存在相关性。

结论

采用ATR-FTIR光谱法分析指甲中的蛋白糖基化可能是一种用于诊断和监测糖尿病的经济实惠的替代技术。由于该检测不消耗试剂,且分析前阶段极为稳定,因此该检测在发展中国家可能特别有用。

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