基于NADH荧光检测的光纤山梨糖醇生物传感器可快速诊断糖尿病并发症<a href="#fn1" title="提供电子补充信息（ESI）。参见DOI：10.1039 / c4an01593b">†</a>,Analyst

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基于NADH荧光检测的光纤山梨糖醇生物传感器可快速诊断糖尿病并发症 †
Analyst ( IF 3.6 ) Pub Date : 2015-08-05 00:00:00 , DOI: 10.1039/c4an01593b
Tomoko Gessei _{1,

2,

3,

4,

5} , Takahiro Arakawa _{2,

3,

5,

6} , Hiroyuki Kudo _{2,

3,

5,

6} , Kohji Mitsubayashi _{2,

3,

4,

5,

6}

Affiliation

已知组织中山梨糖醇的积累会引起微血管糖尿病并发症。在本文中，开发并测试了用于山梨糖醇的光纤生物传感器，该传感器被用作糖尿病并发症的生物标记。该生物传感器使用了来自黄病毒属微生物的山梨糖醇脱氢酶。具有高底物特异性，并通过酶促反应检测到还原的烟酰胺腺嘌呤二核苷酸（NADH）的荧光。紫外线发光二极管（UV-LED）被用作NADH的激发光源。使用分光计或光电倍增管（PMT）检测NADH的荧光。UV-LED和光电检测器使用Y形光纤耦合。在实验中，将带有山梨糖醇脱氢酶固定膜的光纤探头放入装有比色皿的比色皿中，该磷酸盐缓冲液含有烟酰胺腺嘌呤二核苷酸（NAD ⁺）。加入标准山梨糖醇溶液后，测量NADH荧光强度的变化。根据实验评估，使用光谱仪和PMT的山梨糖醇生物传感器系统的校准范围分别为5.0–1000μmolL ^-1和1.0–1000μmolL ^-1。与来自绵羊肝的微生物相比，使用来自黄病毒属微生物的山梨糖醇脱氢酶的山梨糖醇生物传感器系统具有较高的选择性和灵敏度。山梨糖醇生物传感器可用于糖尿病患者的即时护理测试应用或日常医疗保健测试。

"点击查看英文标题和摘要"

A fiber-optic sorbitol biosensor based on NADH fluorescence detection toward rapid diagnosis of diabetic complications†

Accumulation of sorbitol in the tissue is known to cause microvascular diabetic complications. In this paper, a fiber-optic biosensor for sorbitol which is used as a biomarker of diabetic complications was developed and tested. The biosensor used a sorbitol dehydrogenase from microorganisms of the genus Flavimonas with high substrate specificity and detected the fluorescence of reduced nicotinamide adenine dinucleotide (NADH) by the enzymatic reaction. An ultraviolet light emitting diode (UV-LED) was used as the excitation light source of NADH. The fluorescence of NADH was detected using a spectrometer or a photomultiplier tube (PMT). The UV-LED and the photodetector were coupled using a Y-shaped optical fiber. In the experiment, an optical fiber probe with a sorbitol dehydrogenase immobilized membrane was placed in a cuvette filled with a phosphate buffer containing the oxidized form of nicotinamide adenine dinucleotide (NAD⁺). The changes in NADH fluorescence intensity were measured after adding a standard sorbitol solution. According to the experimental assessment, the calibration range of the sorbitol biosensor systems using a spectrometer and a PMT was 5.0–1000 μmol L⁻¹ and 1.0–1000 μmol L⁻¹, respectively. The sorbitol biosensor system using the sorbitol dehydrogenase from microorganisms of the genus Flavimonas has high selectivity and sensitivity compared with that from sheep liver. The sorbitol biosensor allows for point-of-care testing applications or daily health care tests for diabetes patients.

更新日期：2015-08-05

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