检出限
离心机
蛋白质检测
数字聚合酶链反应
色谱法
人血浆
重组DNA
化学
生物医学工程
材料科学
纳米技术
医学
聚合酶链反应
物理
生物化学
基因
核物理学
作者
Zhonghua Tang,Feng Lv,David Eun Reynolds,Shunji Zhang,Shufa Zheng,Jina Ko,Yu Chen,Yongcheng Wang
出处
期刊:Lab on a Chip
[The Royal Society of Chemistry]
日期:2023-01-01
卷期号:23 (12): 2758-2765
被引量:3
摘要
The ability to efficiently detect low-abundance protein biomarkers in tiny blood samples is a significant challenge in clinical and laboratory settings. Currently, high-sensitivity approaches require specialized instrumentation, involve multiple washing steps, and lack the ability to parallelize, preventing their widespread implementation. Herein, we developed a parallelized, wash-free, and ultrasensitive centrifugal droplet digital protein detection (CDPro) technology that achieves a femtomolar limit of detection (LoD) of target proteins with sub-microliters of plasma. The CDPro combines two techniques, namely a centrifugal microdroplet generation device and a digital immuno-PCR assay. Miniaturized centrifugal devices enable emulsification of hundreds of samples within 3 minutes using a common centrifuge. The bead-free digital immuno-PCR assay not only eliminates the need for multistep washing, but also possesses ultra-high detection sensitivity and accuracy. We characterized the performance of CDPro using recombinant interleukins (IL-3 and IL-6) as example targets and reported a LoD of 0.0128 pg mL-1. We also quantified IL-6 from 7 human clinical blood samples using the CDPro with only 0.5 μL plasma, which showed excellent agreement with an existing clinical protein diagnostic system with 25 μL plasma from those samples (R2 = 0.98).
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