Magnetic Optical Microarray Imager for Diagnosing Type of Diabetes in Clinical Blood Serum Samples.
Magnetic Optical Microarray Imager for Diagnosing Type of Diabetes in Clinical Blood Serum Samples.
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DOI：
10.1021/acssensors.5b00273
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发表时间：
2016-04-22
期刊：
影响因子：
8.9
通讯作者：
Krishnan S
中科院分区：
文献类型：
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作者：
Singh V;Rodenbaugh C;Krishnan S
由于全球糖尿病和糖尿病前期疾病及其相关致命并发症的发病率迅速上升，设计新的诊断工具以可靠和直接地测量临床样本中的胰岛素水平势在必行。在这里，我们报告了一种使用表面等离子体共振微阵列成像仪(SPRI)对糖尿病患者样本中的胰岛素水平进行简单而灵敏的直接成像的方法。为了提高灵敏度，我们利用磁性纳米颗粒(MNPs)直接或通过固定在MNPs上的捕获抗体从血清样本中捕获胰岛素。捕获胰岛素的纳米颗粒被允许结合表面胰岛素抗体，以便使用内置SPRI的电荷耦合器件(CCD)从像素强度增加进行检测。我们比较了SPRI免疫阵列在检测不同百分比血清溶液中制备的胰岛素的分析优值。用四参数Logistic模型得到了胰岛素浓度与芯片响应的最佳拟合度，表明胰岛素-纳米颗粒结合物在缓冲胰岛素和血清胰岛素与MNPs结合物中都与表面抗体协同结合。协同效应归因于磁性纳米颗粒结合的胰岛素分子与与表面抗体结合的胰岛素浓度的增加有更大的关联性。这是SPRI免疫阵列的第一份报告，该阵列基于血清基质效应分析和直接和三明治胰岛素测定格式的比较，基于胰岛素水平完成对糖尿病和糖尿病前期疾病的临床诊断。
Due to rapidly rising rates of diabetes and prediabetic conditions worldwide and the associated lethal complications, it is imperative to devise new diagnostic tools that reliably and directly measure insulin levels in clinical samples. Herein, we report a simple and sensitive direct imaging of insulin levels in diabetic patient samples using a surface plasmon resonance microarray imager (SPRi). To enhance sensitivity, we utilized magnetic nanoparticles (MNPs) to capture insulin from serum samples either directly or via a capture antibody immobilized on MNPs. The insulin-captured nanoparticles were allowed to bind surface insulin-antibody for detection from pixel intensity increase using a charge coupled device (CCD) built-in with the SPRi. We have compared the analytical figures-of-merit of the SPRi immunoarray on detecting insulin prepared in various percentages of serum solutions. A four parameter logistic model was used to obtain the best fit of microarray responses with insulin concentration and indicated the cooperative binding of insulin–nanoparticle conjugates to surface antibody in both the buffer insulin and the serum insulin conjugates with MNPs. The cooperativity effect is attributed to the greater association of magnetic nanoparticle-bound insulin molecules with increasing concentration of insulin binding to surface antibody. This is the first report of an SPRi immunoarray to accomplish clinical diagnosis of diabetic and prediabetic conditions based on insulin levels with serum matrix effect analysis and comparison between direct and sandwich insulin assay formats.