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用于疟原虫感染红细胞的高通量无标记寄生虫血症定量和阶段分化

High-throughput and label-free parasitemia quantification and stage differentiation for malaria-infected red blood cells.

作者信息

Yang Xiaonan, Chen Zhuofa, Miao Jun, Cui Liwang, Guan Weihua

机构信息

Department of Electrical Engineering, Pennsylvania State University, University Park 16802, USA; School of Information Engineering, Zhengzhou University, Zhengzhou 450000, China.

Department of Electrical Engineering, Pennsylvania State University, University Park 16802, USA.

出版信息

Biosens Bioelectron. 2017 Dec 15;98:408-414. doi: 10.1016/j.bios.2017.07.019. Epub 2017 Jul 8.

Abstract

This work reports a high throughput and label-free microfluidic cell deformability sensor for quantitative parasitemia measurement and stage determination for Plasmodium falciparum-infected red blood cells (Pf-iRBCs). The sensor relies on differentiating the RBC deformability (a mechanical biomarker) that is highly correlated with the infection status. The cell deformability is measured by evaluating the transit time when each individual RBC squeezes through a microscale constriction (cross-section ~5µm×5µm). More than 30,000 RBCs can be analyzed for parasitemia quantification in under 1min with a throughput ~500 cells/s. Moreover, the device can also differentiate various malaria stages (ring, trophozoite, and schizont stage) due to their varied deformability. Using Pf-iRBCs at 0.1% parasitemia as a testing sample, the microfluidic deformability sensor achieved an excellent sensitivity (94.29%), specificity (86.67%) and accuracy (92.00%) in a blind test, comparable to the gold standard of the blood smear microscopy. As a supplement technology to the microscopy and flow cytometry, the microfluidic deformability sensor would possibly allow for label-free, rapid and cost-effective parasitemia quantification and stage determination for malaria in remote regions.

摘要

这项工作报道了一种用于定量恶性疟原虫感染的红细胞(Pf-iRBCs)的寄生虫血症和确定其阶段的高通量、无标记微流控细胞变形性传感器。该传感器依赖于区分与感染状态高度相关的红细胞变形性(一种机械生物标志物)。通过评估每个红细胞挤过微尺度收缩处(横截面约5μm×5μm)的通过时间来测量细胞变形性。在不到1分钟的时间内,可以分析超过30000个红细胞用于寄生虫血症定量,通量约为500个细胞/秒。此外,由于不同疟疾阶段(环状体、滋养体和裂殖体阶段)的变形性不同,该设备还可以区分它们。以0.1%寄生虫血症的Pf-iRBCs作为测试样本,微流控变形性传感器在盲测中实现了优异的灵敏度(94.29%)、特异性(86.67%)和准确性(92.00%),与血涂片显微镜检查的金标准相当。作为显微镜检查和流式细胞术的补充技术,微流控变形性传感器可能会为偏远地区的疟疾提供无标记、快速且经济高效的寄生虫血症定量和阶段确定方法。

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