4.7 Article

A Point-of-Care Device for Fully Automated, Fast and Sensitive Protein Quantification via qPCR

期刊

BIOSENSORS-BASEL
卷 12, 期 7, 页码 -

出版社

MDPI
DOI: 10.3390/bios12070537

关键词

point-of-care; noise minimisation; qPCR; algorithm; diagnostics; protein quantification; DNA aptamers

资金

  1. ERC Proof of Concept grant [825796]
  2. ERC Synergy Grant [319818]
  3. UK EPSRC [EP/N002474/1]
  4. UK EPSRC Doctoral Training Partnership
  5. European Research Council (ERC) [825796, 319818] Funding Source: European Research Council (ERC)

向作者/读者索取更多资源

This paper presents a fully automated point-of-care device for protein quantification using short-DNA aptamers. The device integrates DNA amplification, sensing, and real-time data processing, and can be used for disease diagnostics and personalized medicine. Additionally, a new algorithm is proposed for automated and real-time protein quantification. An alternative method is also introduced, which utilizes a convolutional neural network algorithm to classify samples and provide clinically relevant advice.
This paper presents a fully automated point-of-care device for protein quantification using short-DNA aptamers, where no manual sample preparation is needed. The device is based on our novel aptamer-based methodology combined with real-time polymerase chain reaction (qPCR), which we employ for very sensitive protein quantification. DNA amplification through qPCR, sensing and real-time data processing are seamlessly integrated into a point-of-care device equipped with a disposable cartridge for automated sample preparation. The system's modular nature allows for easy assembly, adjustment and expansion towards a variety of biomarkers for applications in disease diagnostics and personalised medicine. Alongside the device description, we also present a new algorithm, which we named PeakFluo, to perform automated and real-time quantification of proteins. PeakFluo achieves better linearity than proprietary software from a commercially available qPCR machine, and it allows for early detection of the amplification signal. Additionally, we propose an alternative way to use the proposed device beyond the quantitative reading, which can provide clinically relevant advice. We demonstrate how a convolutional neural network algorithm trained on qPCR images can classify samples into high/low concentration classes. This method can help classify obese patients from their leptin values to optimise weight loss therapies in clinical settings.

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