Bubble-free rapid microfluidic PCR

Lee, Sang Hun; Song, Jihwan; Cho, Byungrae; Hong, SoonGweon; Hoxha, Ori; Kang, Taewook; Kim, Dongchoul; Lee, Luke P.

doi:10.1016/j.bios.2018.10.005

Cited by 56 publications

(46 citation statements)

References 38 publications

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“…Moreover, cortisol is a clinically proven stress biomarker that plays a vital role in the regulation of various physiological processes such as blood pressure, glucose levels, and carbohydrate metabolism [1,2]. Variations or abnormalities in physiological cortisol levels regulate acute and chronic stress responses; they also differentiate between disease states and thus serve as indicators of progression [3][4][5]. For example, excess cortisol levels contribute to the development of Cushing's disease, including its symptoms of obesity, fatigue, and bone fragility [6].…”

Section: Introductionmentioning

confidence: 99%

Ultrasensitive Stress Biomarker Detection Using Polypyrrole Nanotube Coupled to a Field-Effect Transistor

et al. 2020

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Stress biomarkers such as hormones and neurotransmitters in bodily fluids can indicate an individual’s physical and mental state, as well as influence their quality of life and health. Thus, sensitive and rapid detection of stress biomarkers (e.g., cortisol) is important for management of various diseases with harmful symptoms, including post-traumatic stress disorder and depression. Here, we describe rapid and sensitive cortisol detection based on a conducting polymer (CP) nanotube (NT) field-effect transistor (FET) platform. The synthesized polypyrrole (PPy) NT was functionalized with the cortisol antibody immunoglobulin G (IgG) for the sensitive and specific detection of cortisol hormone. The anti-cortisol IgG was covalently attached to a basal plane of PPy NT through an amide bond between the carboxyl group of PPy NT and the amino group of anti-cortisol IgG. The resulting field-effect transistor-type biosensor was utilized to evaluate various cortisol concentrations. Cortisol was sensitively measured to a detection limit of 2.7 × 10−10 M (100 pg/mL), with a dynamic range of 2.7 × 10−10 to 10−7 M; it exhibited rapid responses (<5 s). We believe that our approach can serve as an alternative to time-consuming and labor-intensive health questionnaires; it can also be used for diagnosis of underlying stress-related disorders.

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Section: Introductionmentioning

confidence: 99%

Ultrasensitive Stress Biomarker Detection Using Polypyrrole Nanotube Coupled to a Field-Effect Transistor

et al. 2020

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“…In microfluidic PCR, thermocycling remains an important challenge which at times can be exacerbated by miniaturization and microfluidic implementation 23 , alongside challenges in device integration 23 , 24 . This is particularly challenging in dPCR where homogenous amplification is essential.…”

Section: Introductionmentioning

confidence: 99%

Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells

Hardinge

Baxani

McCloy

et al. 2020

Sci Rep

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Microfluidic droplet generation affords precise, low volume, high throughput opportunities for molecular diagnostics. Isothermal DNA amplification with bioluminescent detection is a fast, low-cost, highly specific molecular diagnostic technique that is triggerable by temperature. Combining loop-mediated isothermal nucleic acid amplification (LAMP) and bioluminescent assay in real time (BART), with droplet microfluidics, should enable high-throughput, low copy, sequence-specific DNA detection by simple light emission. Stable, uniform LAMP–BART droplets are generated with low cost equipment. The composition and scale of these droplets are controllable and the bioluminescent output during DNA amplification can be imaged and quantified. Furthermore these droplets are readily incorporated into encapsulated droplet interface bilayers (eDIBs), or artificial cells, and the bioluminescence tracked in real time for accurate quantification off chip. Microfluidic LAMP–BART droplets with high stability and uniformity of scale coupled with high throughput and low cost generation are suited to digital DNA quantification at low template concentrations and volumes, where multiple measurement partitions are required. The triggerable reaction in the core of eDIBs can be used to study the interrelationship of the droplets with the environment and also used for more complex chemical processing via a self-contained network of droplets, paving the way for smart soft-matter diagnostics.

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“…PDMS, a widely used polymer in microfluidic fabrication, triggers massive gas escape during the heating process. Thin films such as Parylene C, fluorosilane polymer and polyethylene as gas vapor barriers are integrated into the MF chip [36][37][38] . However, the process of depositing these materials is complex and time-consuming.…”

Section: Design Concept For the Simple Operation Mf-lamp Chipmentioning

confidence: 99%

Passively Driven Microfluidic Device with Simple Operation for Quantitative Droplet LAMP Sssay for Point-of-Care Analysis

Lin

2021

Preprint

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Since polymerase chain reaction (PCR) has become a vital tool for disease diagnosis, the development of precise applied PCR technologies in point-of care testing (POCT) has become more significant. The microfluidic-based PCR platform offers a great opportunity for on-site diagnosis efficiency, and the system is aimed at user-friendly access. Herein, we demonstrate a microfluidic system with simple operation that provides reliable nucleic acid results from 18 uniform droplets via LAMP qPCR detection. By using only micropipette regulation, users are able to control the nanoliter scale of the droplets in this valve-free and pump-free microfluidic (MF) chip. Based on the oil enclosure method and impermeable fabrication, we successfully preserved the reagent inside the microfluidic system, which significantly reduced the fluid loss and condensation. The relative standard deviation (RSD) of the fluorescence intensity between the droplets and during the heating process was <5% and 2.0%, respectively. Additionally, for different nucleic acid detection methods, the MF-LAMP chip in this study showed good applicability to both genome detection and gene expression analysis.

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Bubble-free rapid microfluidic PCR

Cited by 56 publications

References 38 publications

Ultrasensitive Stress Biomarker Detection Using Polypyrrole Nanotube Coupled to a Field-Effect Transistor

Ultrasensitive Stress Biomarker Detection Using Polypyrrole Nanotube Coupled to a Field-Effect Transistor

Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells

Passively Driven Microfluidic Device with Simple Operation for Quantitative Droplet LAMP Sssay for Point-of-Care Analysis

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