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用于即时检测血清谷氨酸草酰乙酸转氨酶(SGOT)的 µPAD 的开发。

Development of a µPAD for the point-of-care testing of serum glutamic oxaloacetic transaminase (SGOT).

机构信息

Department of Chemistry, Amrita School of Physical Sciences Coimbatore, Amrita Vishwa Vidyapeetham, Coimbatore, 641112, India.

Amrita Biosensor Research Lab, Amrita School of Engineering Coimbatore, Amrita Vishwa Vidyapeetham, Coimbatore, 641112, India.

出版信息

Mikrochim Acta. 2024 Sep 25;191(10):619. doi: 10.1007/s00604-024-06678-6.

Abstract

A wax-patterned paper analytical device (µPAD) has been developed for point-of-care colourimetric testing of serum glutamic oxaloacetic transaminase (SGOT). The detection method was based on the transamination reaction of aspartate with α-ketoglutarate, leading to the formation of oxaloacetate which reacts with the reagent Fast Blue BB salt and forms a cavern pink colour. The intensity of the cavern pink colour grows as the concentration of SGOT increases. UV-visible spectroscopy was utilized to optimize reaction conditions, and the optimized reagents were dropped onto the wax-patterned paper. The coloured PADs, after the addition of SGOT, have been photographed, and a colour band has been generated to correlate the SGOT concentration visually. The images were used to calculate the intensity values using ImageJ software, which inturn was used to calculate the SGOT concentration. The PADs were also tested with serum samples, and SGOT spiked serum samples. The PAD could detect the SGOT concentration ranging from 5 to 200 U/L. The analysis yielded highly accurate results with less than 6% relative error compared to the clinical sample. This colourimetric test demonstrated exceptional selectivity in the presence of other biomolecules in the blood serum, with a detection limit of 2.77 U/L and a limit of quantification of 9.25 U/L. Additionally, a plasma separation membrane was integrated with the PAD to directly test SGOT from finger-prick blood samples.

摘要

已开发出一种蜡模纸分析装置(µPAD),用于即时检测血清谷氨酸草酰乙酸转氨酶(SGOT)的比色分析。该检测方法基于天冬氨酸与α-酮戊二酸的转氨基反应,形成草酰乙酸,然后与试剂 Fast Blue BB 盐反应,形成洞穴粉红色。洞穴粉红色的强度随着 SGOT 浓度的增加而增加。利用紫外可见光谱优化反应条件,并将优化后的试剂滴加到蜡模纸上。加入 SGOT 后,对显色的 PAD 进行拍照,并生成色带,用于目视法关联 SGOT 浓度。使用 ImageJ 软件对图像进行强度值计算,进而计算 SGOT 浓度。还使用血清样品和添加 SGOT 的血清样品对 PAD 进行了测试。PAD 可以检测 5 至 200 U/L 范围内的 SGOT 浓度。与临床样本相比,该分析的相对误差小于 6%,结果高度准确。与血液中的其他生物分子相比,这种比色试验在存在其他生物分子时表现出出色的选择性,检测限为 2.77 U/L,定量限为 9.25 U/L。此外,还将血浆分离膜与 PAD 集成,可直接从指尖血样中测试 SGOT。

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