阵列纸芯片比色法检测碱性磷酸酶

引用本文: 陈熙, 陈锦, 张慧妍, 汪付兵, 王方方, 吉邢虎, 何治柯. 阵列纸芯片比色法检测碱性磷酸酶[J]. 分析化学, 2016, 44(4): 591-596. doi: 10.11895/j.issn.0253-3820.160140 shu

Citation: CHEN Xi, CHEN Jin, ZHANG Hui-Yan, WANG Fu-Bing, WANG Fang-Fang, JI Xing-Hu, HE Zhi-Ke. Colorimetic Detection of Alkaline Phosphatase on Microfluidic Paper-based Analysis Devices[J]. Chinese Journal of Analytical Chemistry, 2016, 44(4): 591-596. doi: 10.11895/j.issn.0253-3820.160140 shu

阵列纸芯片比色法检测碱性磷酸酶

陈熙 ^1, ,
陈锦 ^1, ,
张慧妍 ^1, ,
汪付兵 ^2, ,
王方方 ^1, ,
吉邢虎 ^{1,
,} ,
何治柯 ^1,

1.
¹ 生物医学分析化学教育部重点实验室, 武汉大学化学与分子科学学院, 武汉 430072;
2.
² 武汉大学中南医院检验科, 武汉 430071

通讯作者: 吉邢虎

基金项目:
本文系国家自然科学基金(No.21205089) (No.21205089)

教育部高等学校博士学科点专项科研基金(No.20120141120036)资助项目 (No.20120141120036)

摘要: 采用5-溴-4-氯-3-吲哚磷酸盐(BCIP)/氯化硝基四氮唑蓝(NBT)显色体系,构建了阵列纸芯片比色检测碱性磷酸酶(ALP)的方法。首先,借助烘干处理方式在光刻法制备的阵列纸芯片微孔中固定显色试剂,然后加入ALP进行显色反应,最后,采用凝胶成像仪和普通照相机成像,读取显色强度(灰度值)进行比色检测。详细考察了显色条件对检测结果的影响,探讨了人血清白蛋白对ALP检测的增色效应,在最佳实验条件下,ALP检测的线性范围为1.5~20 U/L,检出限(3σ)为0.78 U/L(n=18),比文献报道中纸芯片上检测ALP方法的检出限低约两个数量级。本方法成功用于实际血清样品检测,测定结果与临床值一致。在此基础上,构建了双色阵列纸芯片,通过颜色的变化实现了ALP的可视化半定量检测。

关键词:

English

Colorimetic Detection of Alkaline Phosphatase on Microfluidic Paper-based Analysis Devices

1.
¹ Key Laboratory of Analytical Chemistry for Biology and Medicine, Ministry of Education, College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, China;
2.
² Department of Laboratory Medicine, Zhongnan Hospital of Wuhan University, Wuhan 430071, China

Corresponding author: 吉邢虎

Received Date: 25 February 2016
Fund Project:
本文系国家自然科学基金(No.21205089)

教育部高等学校博士学科点专项科研基金(No.20120141120036)资助项目

Abstract: A low-cost, simple and sensitive colorimetric detection method of alkaline phosphatase(ALP) has been developed on microfluidic paper-based analysis devices. In a typical colorimetric detection, 5-bromo-4-chloro-3-indolyl phosphate disodium salt(BCIP) and nitro blue tetrazolium(NBT) were firstly added onto the circle array zones of paper devices. Then ALP solutions were spotted to the array zones to perform the colorimetric reaction. The color results were recorded by both Gel Documentation systems and a common camera, and analyzed with Quantity One software. All the reaction conditions were aptimized. Under the optimal conditions, the colorimetric intensity showed a linear correlation to the concentration of ALP in the range of 1.5 to 20 U/L with the limit of detection(LOD) of 0.78 U/L(3σ), which was about two orders of magnitude lower than that of the reported method on μPADs. Besides, it could be applied in spiked real sample analysis with a satisfactory result. In addition, a dual-color array-based paper strip has been fabricated for the semiquantitative detection of ALP. The approximate activity of ALP could be simply distinguished by observing the color change with naked eyes.

Key words:

Microfluidic paper-based analysis devices
/ Colorimetric detection
/ Alkaline phosphatase

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

阵列纸芯片比色法检测碱性磷酸酶

通讯作者: 吉邢虎

English

Colorimetic Detection of Alkaline Phosphatase on Microfluidic Paper-based Analysis Devices

Corresponding author: 吉邢虎

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

阵列纸芯片比色法检测碱性磷酸酶

通讯作者: 吉邢虎

English

Colorimetic Detection of Alkaline Phosphatase on Microfluidic Paper-based Analysis Devices

Corresponding author: 吉邢虎

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

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CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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