阵列纸芯片比色法检测碱性磷酸酶

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（3#）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...

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Bibliographic Details
Published in分析化学 Vol. 44; no. 4; pp. 591 - 596
Main Author 陈熙 陈锦 张慧妍 汪付兵 王方方 吉邢虎 何治柯
Format Journal Article
LanguageChinese
Published 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%武汉大学中南医院检验科,武汉,430071 2016
Subjects
比色法
碱性磷酸酶
阵列纸芯片
Alkaline phosphatase
阵列纸芯片
比色法
碱性磷酸酶
Microfluidic paper-based analysis devices
Colorimetric detection
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ISSN0253-3820
DOI10.11895/j.issn.0253-3820.160140

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Summary: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（3#）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。
Bibliography:Microfluidic paper-based analysis devices; Colorimetric detection; Alkaline phosphatase
CHEN Xi;CHEN Jin;ZHANG Hui-Yan;WANG Fu-Bing;WANG Fang-Fang;JI Xing-Hu;HE Zhi-Ke; 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 （ Department of Laboratory Medicine, Zhongnan Hospital of Wuhan University, Wuhan 430071, China）
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
ISSN:0253-3820
DOI:10.11895/j.issn.0253-3820.160140