Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, Taiwan.
Biosens Bioelectron. 2015 Jun 15;68:397-403. doi: 10.1016/j.bios.2015.01.027. Epub 2015 Jan 13.
Blood glycated hemoglobin (HbA1c), reflecting the average blood glucose level in the proceeding 2-3 months, is recommended for screening/diagnosing and patient management of diabetes. However, accurate measurement of the HbA1c level at the point of care is hampered by costly, large-scale instruments (such as high-performance liquid chromatography) or reagent instability of classical immunologic methods, which involve antibody-based immunoturbidimetry. In this work, an integrated microfluidic system using aptamer-based testing to measure HbA1c in blood samples is therefore presented. This measuring system used nucleic-acid aptamers that exhibited high sensitivity and high specificity for hemoglobin and HbA1c to perform a stable and robust testing. The compact microfluidic system consumed less samples and reagents and significantly shortened the detection time. Combining the advantages of microfluidics and aptamers, this integrated microsystem presents a promising tool for accurate and point-of-case HbA1c detection. To demonstrate its clinical utility, whole blood samples with clinically-relevant concentrations of HbA1c and Hb were automatically measured on the integrated microfluidic system. Experimental data showed that the developed aptamer-based microfluidic system is capable of detecting HbA1c and Hb with a good linear response. The entire process was completed within 25 min. The aptamer-antibody on-chip sandwich immunoassay may be further refined to allow diabetes screening and diagnosis at lower cost and earlier phase to minimize the risk of diabetic complications.
糖化血红蛋白（HbA1c）反映了过去 2-3 个月的平均血糖水平，推荐用于糖尿病的筛查/诊断和患者管理。然而，在即时护理点准确测量 HbA1c 水平受到昂贵的大型仪器（如高效液相色谱法）或经典免疫方法的试剂不稳定性的限制，这些方法涉及基于抗体的免疫比浊法。因此，本工作提出了一种使用适体进行基于检测的集成微流控系统来测量血液样本中的 HbA1c。该测量系统使用了对血红蛋白和 HbA1c 表现出高灵敏度和高特异性的核酸适体，以进行稳定和强大的检测。紧凑的微流体系统消耗更少的样本和试剂，并大大缩短了检测时间。该集成微系统结合了微流控和适体的优势，为准确和即时的 HbA1c 检测提供了一种有前途的工具。为了证明其临床实用性，我们在集成微流控系统上自动测量了具有临床相关 HbA1c 和 Hb 浓度的全血样本。实验数据表明，所开发的基于适体的微流控系统能够以良好的线性响应检测 HbA1c 和 Hb。整个过程在 25 分钟内完成。基于适体-抗体的芯片夹心免疫分析可以进一步完善，以允许以更低的成本和更早的阶段进行糖尿病筛查和诊断，从而最大程度地降低糖尿病并发症的风险。
