Point-of-care diagnostics for noncommunicable diseases using synthetic urinary biomarkers and paper microfluidics
Point-of-care diagnostics for noncommunicable diseases using synthetic urinary biomarkers and paper microfluidics
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DOI：
10.1073/pnas.1314651111
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发表时间：
2014-03-11
影响因子：
11.1
通讯作者：
Bhatia, Sangeeta N.
中科院分区：
文献类型：
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作者：
Warren, Andrew D.;Kwong, Gabriel A.;Bhatia, Sangeeta N.
由于非传染性疾病现在构成全球死亡率的大部分，因此越来越需要低成本、非侵入性的方法来诊断和治疗这类疾病，特别是在资源有限的情况下。分子生物标志物与低成本的即时检测相结合，构成了诊断非传染性疾病的潜在解决方案，但缺乏天然存在的预测性标志物限制了这种方法。在这里，我们描述了外源性药物的设计，作为非传染性疾病的合成生物标志物，通过产生尿液信号，可以量化的配套纸测试。这些合成生物标志物由纳米颗粒组成，通过蛋白酶敏感的肽底物与配体编码的报告分子缀合。在递送时，纳米颗粒被动地靶向患病部位，例如实体瘤或血凝块，其中上调的蛋白酶切割肽底物并释放被清除到尿液中的报告物。记者被设计用于检测夹心免疫测定，我们证明他们的定量直接从未经修改的尿液，此外，捕获抗体的特异性允许探针在体内多重和定量同时通过ELISA或纸侧流试验（LFA）。我们定制了针对结直肠癌（一种代表性实体瘤）和血栓形成（一种常见的心血管疾病）的合成生物标志物，并通过纸诊断证明了这些疾病在小鼠模型中的尿液检测。LFA和可注射的合成生物标志物可以针对多种疾病量身定制，共同形成了一个通用的非传染性疾病诊断平台，可以在几乎任何情况下应用，而无需昂贵的设备或训练有素的医务人员。
With noncommunicable diseases (NCDs) now constituting the majority of global mortality, there is a growing need for low-cost, noninvasive methods to diagnose and treat this class of diseases, especially in resource-limited settings. Molecular biomarkers combined with low-cost point-of-care assays constitute a potential solution for diagnosing NCDs, but the dearth of naturally occurring, predictive markers limits this approach. Here, we describe the design of exogenous agents that serve as synthetic biomarkers for NCDs by producing urinary signals that can be quantified by a companion paper test. These synthetic biomarkers are composed of nanoparticles conjugated to ligand-encoded reporters via protease-sensitive peptide substrates. Upon delivery, the nanoparticles passively target diseased sites, such as solid tumors or blood clots, where up-regulated proteases cleave the peptide substrates and release reporters that are cleared into urine. The reporters are engineered for detection by sandwich immunoassays, and we demonstrate their quantification directly from unmodified urine; furthermore, capture antibody specificity allows the probes to be multiplexed in vivo and quantified simultaneously by ELISA or paper lateral flow assay (LFA). We tailor synthetic biomarkers specific to colorectal cancer, a representative solid tumor, and thrombosis, a common cardiovascular disorder, and demonstrate urinary detection of these diseases in mouse models by paper diagnostic. Together, the LFA and injectable synthetic biomarkers, which could be tailored for multiple diseases, form a generalized diagnostic platform for NCDs that can be applied in almost any setting without expensive equipment or trained medical personnel.