Quantitative and multiplex microRNA assays from unprocessed cells in isolated nanoliter well arrays†,Lab on a Chip

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Quantitative and multiplex microRNA assays from unprocessed cells in isolated nanoliter well arrays†
Lab on a Chip ( IF 6.1 ) Pub Date : 2018-07-06 00:00:00 , DOI: 10.1039/c8lc00498f
Augusto M. Tentori _{1,

2,

3,

4} , Maxwell B. Nagarajan _{1,

2,

3,

4} , Jae Jung Kim _{1,

2,

3,

4} , Wen Cai Zhang _{4,

5,

6,

7} , Frank J. Slack _{4,

5,

6,

7} , Patrick S. Doyle _{1,

2,

3,

4}

Affiliation

MicroRNAs (miRNAs) have recently emerged as promising biomarkers for the profiling of diseases. Translation of miRNA biomarkers to clinical practice, however, remains a challenge due to the lack of analysis platforms for sensitive, quantitative, and multiplex miRNA assays that have simple and robust workflows suitable for translation. The platform we present here utilizes functionalized hydrogel posts contained within isolated nanoliter well reactors for quantitative and multiplex assays directly from unprocessed cell samples without the need of prior nucleic acid extraction. Simultaneous reactor isolation and delivery of miRNA extraction reagents is achieved by sealing an array of wells containing the functionalized hydrogel posts and cells against another array of wells containing lysis and extraction reagents. The nanoliter well array platform features >100× better sensitivity compared to previous technology utilizing hydrogel particles without relying on signal amplification and enables >100 parallel assays in a single device. These advances provided by this platform lay the groundwork for translatable and robust analysis technologies for miRNA expression profiling in samples with small populations of cells and in precious, material-limited samples.

中文翻译：

分离的纳升孔阵列中未处理细胞的定量和多重microRNA测定†

微小RNA（miRNA）最近已成为有前途的疾病分析生物标志物。然而，由于缺乏用于敏感，定量和多重miRNA分析的分析平台，而这些平台具有适合翻译的简单而强大的工作流程，因此将miRNA生物标记物翻译到临床实践仍然是一个挑战。我们在此展示的平台利用分离的纳升孔反应器中包含的功能化水凝胶柱，直接从未经处理的细胞样品中进行定量和多重测定，而无需事先进行核酸提取。通过将一系列包含功能化水凝胶柱和细胞的孔与另一组含有裂解和提取试剂的孔密封，可以实现反应器同时分离和递送miRNA提取试剂。与以前的利用水凝胶颗粒而不依赖信号放大的技术相比，纳升孔阵列平台的灵敏度提高了100倍以上，并且可以在单个设备中进行100多个平行测定。该平台提供的这些进展为可翻译和强大的分析技术奠定了基础，这些技术可用于在细胞数量少的样品中以及在材料有限的珍贵样品中进行miRNA表达谱分析。

更新日期：2018-07-06

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