Department of Mechanical Engineering, Indian Institute of Technology (BHU), Varanasi-221005, India.
Department of Electronics Engineering, Indian Institute of Technology (BHU), Varanasi-221005, India.
Anal Methods. 2024 Nov 14;16(44):7546-7556. doi: 10.1039/d4ay01476f.
Creatinine, an important biomarker for renal functions, is often conventionally estimated using the gold standard Jaffe reaction from blood, which involves the usage of a spectrophotometric measurement, thus restricting its utilization scope in point-of-care settings. Here, we report the development of a method for the single-step quantification of creatinine from whole blood using a paper-based microfluidic device. Our platform uses Whatman filter paper integrated with an LF1 membrane. The on-chip separation of blood plasma is achieved through the LF1 membrane, while the Whatman component of the device contains the embedded reagents for the Jaffe reaction. The combination of two different grades of paper enables a single-step quantification of creatinine as the separated blood plasma traverses to the reaction zone through capillary imbibition. Colorimetric readouts were quantified using an RGB sensor instead of a smartphone, which is highly platform dependent and incurs a relatively higher cost compared to the other components in typical point-of care (POC) devices. Our sensor was integrated within a 3D box, thereby making the detection virtually instrument free and perfectly suited for POC settings. The limit of detection (LOD) of our device was 0.219 mg dL, which falls within the lower range of physiological values. The coefficient of determination () for the linearity and median accuracy were 0.978 and 94.047%, respectively. The relative standard deviation (RSD) for precision measurements remained below 5% for the developed protocol. Furthermore, we validated the performance of our device with 35 clinical samples in laboratory settings against the gold standard measurements. Our Bland-Altman plot as well as -test and chi-square test results clearly confirmed the validity of our device within a 95% confidence interval.
肌酸酐是肾功能的一个重要生物标志物,通常使用传统的血液 Jaffe 反应进行常规估计,该反应涉及分光光度测量,因此限制了其在即时护理环境中的应用范围。在这里,我们报告了一种使用基于纸张的微流控设备从全血中单步定量肌酸酐的方法。我们的平台使用与 LF1 膜集成的 Whatman 滤纸。通过 LF1 膜实现了血浆的片上分离,而设备的 Whatman 部分包含 Jaffe 反应的嵌入式试剂。两种不同等级的纸张的组合使得可以通过毛细吸入使分离的血浆穿过反应区来进行肌酸酐的单步定量。比色读数使用 RGB 传感器而不是智能手机进行量化,与典型的即时护理 (POC) 设备中的其他组件相比,智能手机高度依赖平台且成本相对较高。我们的传感器集成在 3D 盒内,从而使检测几乎无需仪器,非常适合 POC 设置。我们的设备的检测限 (LOD) 为 0.219mg/dL,属于生理值的较低范围。线性和中位数准确度的决定系数 () 分别为 0.978 和 94.047%。对于所开发的方案,精密度测量的相对标准偏差 (RSD) 保持在 5%以下。此外,我们在实验室环境中使用 35 个临床样本针对黄金标准测量值验证了我们设备的性能。我们的 Bland-Altman 图以及 -检验和卡方检验结果清楚地证实了我们的设备在 95%置信区间内的有效性。