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无电源片上酶免疫分析检测血清前列腺特异抗原(PSA)。

Power-free chip enzyme immunoassay for detection of prostate specific antigen (PSA) in serum.

机构信息

Yousef Jameel Science and Technology Research Center (YJ-STRC), The American University in Cairo, AUC Ave., PO Box 74, New Cairo 11835, Egypt.

出版信息

Biosens Bioelectron. 2013 Nov 15;49:478-84. doi: 10.1016/j.bios.2013.05.058. Epub 2013 Jun 11.

DOI:10.1016/j.bios.2013.05.058
PMID:23811482
Abstract

A power-free, portable "Chip EIA" was designed to render the popular Enzyme Linked Immunosorbent Assay (ELISA) more suitable for point-of-care testing. A number of microfluidic platforms have enabled miniaturization of the conventional microtitre plate ELISA, however, they require external pumping systems, valves, and electric power supply. The Chip EIA platform has eliminated the need for pumps and valves through utilizing a simple permanent magnet and magnetic nanoparticles. The magnetic nanoparticles act as solid support to capture the target and are then moved through chambers harboring different reagents necessary to perform a sandwich ELISA. The use of magnetic nanoparticles increases the volume-to-surface ratio reducing the assay time to 30 min. Changing the color of horseradish peroxidase (HRP) substrate to green indicates a positive result. In addition, a quantitative read-out was obtained through the use of cellphone camera imaging and analyzing the images using Matlab®. Cell phones, including smart ones, are readily available almost everywhere. The Chip EIA device was used to assay total prostate specific antigen (tPSA) in 19 serum samples. The PSA Chip EIA was tested for accuracy, precision, repeatability, and the results were correlated to the commercial Beckman Colter, Hybritech immunoassay® for determination of tPSA in serum samples with a Pearson correlation coefficient (R(2)=0.96). The lower detection limit of the PSA Chip EIA was 3.2 ng/mL. The assay has 88.9% recovery and good reproducibility (% CV of 6.5). We conclude that the developed Chip EIA can be used for detection of protein biomarkers in biological specimens.

摘要

一种无电源、便携式的“芯片 EIA”被设计用来使流行的酶联免疫吸附测定(ELISA)更适合于即时检测。许多微流控平台已经实现了传统微量滴定板 ELISA 的微型化,然而,它们需要外部泵送系统、阀门和电源。Chip EIA 平台通过利用简单的永磁体和磁性纳米粒子消除了对泵和阀门的需求。磁性纳米粒子用作捕获靶标的固体支撑物,然后通过容纳执行夹心 ELISA 所需的不同试剂的腔室移动。使用磁性纳米粒子增加了体积与表面积的比率,从而将测定时间缩短至 30 分钟。将辣根过氧化物酶(HRP)底物的颜色变为绿色表示阳性结果。此外,通过使用手机摄像头成像并使用 Matlab®分析图像,可以获得定量读数。手机,包括智能手机,几乎无处不在都很容易获得。该 Chip EIA 设备用于检测 19 份血清样本中的总前列腺特异性抗原(tPSA)。对 PSA Chip EIA 的准确性、精密度、重复性进行了测试,并将结果与商用 Beckman Colter、Hybritech 免疫测定®进行了相关性分析,用于确定血清样本中的 tPSA,皮尔逊相关系数(R(2)=0.96)。PSA Chip EIA 的检测下限为 3.2ng/mL。该测定法的回收率为 88.9%,重现性良好(%CV 为 6.5)。我们得出结论,开发的 Chip EIA 可用于检测生物样本中的蛋白质生物标志物。

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