Deep Learning-Assisted Droplet Digital PCR for Quantitative Detection of Human Coronavirus

Lee, Young Suh; Choi, Ji Wook; Kang, Taewook; Chung, Bong Geun

doi:10.1007/s13206-023-00095-2

Cited by 8 publications

(2 citation statements)

References 38 publications

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“…Limited availability of testing kits, reagents, and laboratory capacity impedes mass testing efforts, leading to delays in identifying positive cases and potential underreporting. Researchers and healthcare professionals have thus explored innovative Biosensors 2024, 14, 91 2 of 9 approaches, such as point-of-care testing, to expedite diagnoses and reach underserved populations [10][11][12][13].…”

Section: Introductionmentioning

confidence: 99%

Dual-Mode Graphene Field-Effect Transistor Biosensor with Isothermal Nucleic Acid Amplification

Kim,

Schuck,

Park

et al. 2024

Biosensors

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Despite a substantial increase in testing facilities during the pandemic, access remains a major obstacle, particularly in low-resource and remote areas. This constraint emphasizes the need for high-throughput potential point-of-care diagnostic tools in environments with limited resources. Loop-mediated isothermal amplification (LAMP) is a promising technique, but improvements in sensitivity are needed for accurate detection, especially in scenarios where the virus is present in low quantities. To achieve this objective, we present a highly sensitive detection approach of a dual-mode graphene-based field-effect transistor (G-FET) biosensor with LAMP. The G-FET biosensor, which has a transparent graphene microelectrode array on a glass substrate, detects LAMP products in less than 30 min using both observable color changes and Dirac point voltage measurements, even in samples with low viral concentrations. This dual-mode G-FET biosensor emerges as a potential alternative to conventional RT-PCR for severe acute respiratory syndrome-associated coronavirus (SARS-CoV)-2 detection or point-of-care testing, particularly in resource-constrained scenarios such as developing countries. Moreover, its capacity for colorimetric detection with the naked eye enhances its applicability in diverse settings.

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Section: Introductionmentioning

confidence: 99%

Dual-Mode Graphene Field-Effect Transistor Biosensor with Isothermal Nucleic Acid Amplification

Kim,

Schuck,

Park

et al. 2024

Biosensors

View full text Add to dashboard Cite

show abstract

“…This feature allows for direct and independent DNA quantification, providing more precise and reproducible data with a strong anti-interference ability [8]. Furthermore, ddPCR has proven to be an effective method for detecting low-abundance targets [9], which holds great potential in pathogen detection applications [5,[10][11][12][13]. The emergence of ddPCR has greatly improved the accuracy and sensitivity of nucleic acid detection.…”

Section: Introductionmentioning

confidence: 99%

An Integrated ddPCR Lab-on-a-Disc Device for Rapid Screening of Infectious Diseases

Zhang,

Cui,

Wang

et al. 2023

Biosensors

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Digital droplet PCR (ddPCR) is a powerful amplification technique for absolute quantification of viral nucleic acids. Although commercial ddPCR devices are effective in the lab bench tests, they cannot meet current urgent requirements for on-site and rapid screening for patients. Here, we have developed a portable and fully integrated lab-on-a-disc (LOAD) device for quantitively screening infectious disease agents. Our designed LOAD device has integrated (i) microfluidics chips, (ii) a transparent circulating oil-based heat exchanger, and (iii) an on-disc transmitted-light fluorescent imaging system into one compact and portable box. Thus, droplet generation, PCR thermocycling, and analysis can be achieved in a single LOAD device. This feature is a significant attribute for the current clinical application of disease screening. For this custom-built ddPCR setup, we have first demonstrated the loading and ddPCR amplification ability by using influenza A virus-specific DNA fragments with different concentrations (diluted from the original concentration to 107 times), followed by analyzing the droplets with an external fluorescence microscope as a standard calibration test. The measured DNA concentration is linearly related to the gradient–dilution factor, which validated the precise quantification for the samples. In addition to the calibration tests using DNA fragments, we also employed this ddPCR-LOAD device for clinical samples with different viruses. Infectious samples containing five different viruses, including influenza A virus (IAV), respiratory syncytial virus (RSV), varicella zoster virus (VZV), Zika virus (ZIKV), and adenovirus (ADV), were injected into the device, followed by analyzing the droplets with an external fluorescence microscope with the lowest detected concentration of 20.24 copies/µL. Finally, we demonstrated the proof-of-concept detection of clinical samples of IAV using the on-disc fluorescence imaging system in our fully integrated device, which proves the capability of this device in clinical sample detection. We anticipate that this integrated ddPCR-LOAD device will become a flexible tool for on-site disease detection.

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Imaging‐Based Efficacy Evaluation of Cancer Immunotherapy in Engineered Tumor Platforms and Tumor Organoids

Kim,

Yun,

Doh

et al. 2024

Adv Healthcare Materials

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Cancer immunotherapy is used to treat tumors by modulating the immune system. Although the anticancer efficacy of cancer immunotherapy has been evaluated prior to clinical trials, conventional in vivo animal and endpoint models inadequately replicate the intricate process of tumor elimination and reflect human‐specific immune systems. Therefore, more sophisticated models that mimic the complex tumor‐immune microenvironment must be employed to assess the effectiveness of immunotherapy. Additionally, using real‐time imaging technology, a step‐by‐step evaluation can be applied, allowing for a more precise assessment of treatment efficacy. Here, we present an overview of the various imaging‐based evaluation platforms recently developed for cancer immunotherapeutic applications. Specifically, we discuss a fundamental technique for stably observing immune cell‐based tumor cell killing using direct imaging, a microwell that reproduces a confined space for spatial observation, a droplet assay that facilitates cell‐cell interactions, and a three‐dimensional microphysiological system that reconstructs the vascular environment. Furthermore, we suggest that future evaluation platforms pursue more human‐like immune systems.This article is protected by copyright. All rights reserved

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Deep Learning-Assisted Droplet Digital PCR for Quantitative Detection of Human Coronavirus

Cited by 8 publications

References 38 publications

Dual-Mode Graphene Field-Effect Transistor Biosensor with Isothermal Nucleic Acid Amplification

Dual-Mode Graphene Field-Effect Transistor Biosensor with Isothermal Nucleic Acid Amplification

An Integrated ddPCR Lab-on-a-Disc Device for Rapid Screening of Infectious Diseases

Imaging‐Based Efficacy Evaluation of Cancer Immunotherapy in Engineered Tumor Platforms and Tumor Organoids

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