Multi-reagents dispensing centrifugal microfluidics for point-of-care testing
Multi-reagents dispensing centrifugal microfluidics for point-of-care testing
复制标题
用于即时检测的多试剂分配离心微流体
DOI：
10.1016/j.bios.2022.114130
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发表时间：
2022-03-01
影响因子：
12.6
通讯作者：
Liu, Bi-Feng
中科院分区：
文献类型：
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作者：
Xiao, Yujin;Li, Shunji;Liu, Bi-Feng
在资源有限的环境中，床旁检测（POCT）在公共卫生监测方面显示出巨大的优势。然而，开发具有多种试剂和样品的自动化和准确定量分配的POCT工具是具有挑战性的。在这里，我们展示了一种新型的多试剂分配离心微流体（MDCM），允许快速和自动化的多个试剂和样品的高通量和准确性分配。MDCM设计有多个等分单元，疏水阀位于不同径向位置。同时加载所有试剂和样品，通过低速离心平行分配，然后通过高速离心依次引入反应室。文中介绍了两种MDCM芯片，包括均匀浓度发生器和梯度浓度发生器。独立反应室间的浓度变异系数（CV）低于0.56%，理论定量浓度梯度与实际浓度梯度呈强相关（R-2 = 0.9938）。我们已成功地将MDCM应用于多种感染性病原体的环介导等温扩增（LAMP）核酸检测和硫酸卡那霉素对大肠杆菌的药敏试验（AST）。杆菌为了进一步扩展应用，MDCM还被应用于具有在线校准的二喹啉甲酸（BCA）蛋白质测定，将检测时间从2小时减少到10分钟，试剂消耗减少了20倍。这些结果表明，MDCM是一个很有潜力的POCT平台。
Point-of-care testing (POCT) has shown great advantages for public health monitoring in resource-limited set-tings. However, developing of POCT tools with automated and accurate quantitative dispensing of multiple reagents and samples is challenging. Here, we demonstrate a novel multi-reagents dispensing centrifugal microfluidics (MDCM) that allows rapid and automated dispensing of multiple reagents and samples with high throughput and accuracy. The MDCM was designed with multiple aliquoting units with the hydrophobic valve at different radial positions. All reagents and samples were loaded simultaneously, dispensed in parallel by centrifugation at low speed, and then introduced into the reaction chamber sequentially by centrifugation at high speed. Two MDCM chips are demonstrated, including a uniform concentration generator and a gradient con-centration generator. The concentration coefficient of variation (CV) among the independent reaction chambers was lower than 0.56%, and the theoretical quantitative concentration gradient was strongly correlated with the actual concentration gradient (R-2 = 0.9938). We have successfully applied the MDCM to loop-mediated isothermal amplification (LAMP)-based nucleic acid detection for multiple infectious pathogens and antimi-crobial susceptibility testing (AST) for kanamycin sulfate against E. coli. To further extend the applications, the MDCM has also been applied to bicinchoninic acid (BCA) protein assays with online calibration, reducing the detection time from 2 h to 10 min with a twenty-fold reduction in reagent consumption. These results indicated that the MDCM is a high potential platform for POCT.