Digital polymerase chain reaction technology – recent advances and future perspectives

Sreejith, Kamalalayam Rajan; Ooi, Chin Hong; Jin, Jing; Dao, Dzung Viet; Nguyen, Nam‐Trung

doi:10.1039/c8lc00990b

Cited by 107 publications

(78 citation statements)

References 149 publications

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“…-stationary chamber PCR [28] is also called "time domain PCR" [29]. A defined volume of sample positioned in a chamber [5] or sealed in a droplet [30] is heated and cooled, either actively using an external element, or passively by simply switching the heater off with a stationary sample. Different heating methods are developed for a fast process, including infrared-mediated [31], plasmonassisted heating [32], and ultrafast with active heating and passive cooling, achieving 40 cycles PCR in <6 min [33].…”

Section: Temperature Cycling Amplificationmentioning

confidence: 99%

“…With an advent of microfluidics methods over the past 25 years, the PCR technique was one of the first utilizing this technology [2]. Since then, the microfluidics were extensively exploited for PCR applications in their conventional [3], quantitative [4], and digital form [5,6], including different POC assays [7,8]. Integration of a conventional PCR with pre-or post-processing samples is difficult, as it requires specially developed hybridization probes, which can pass the nucleic acid (NA) denaturation steps with a temperature of z95 C [9].…”

Section: Introductionmentioning

confidence: 99%

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LAMP-on-a-chip: Revising microfluidic platforms for loop-mediated DNA amplification

Zhang

Fohlerová

et al. 2019

TrAC Trends in Analytical Chemistry

181

126

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a b s t r a c tNucleic acid amplification for the detection of infectious diseases, food pathogens, or assessment of genetic disorders require a laboratory setting with specialized equipment and technical expertise. Isothermal deoxyribonucleic acid amplification methods, such as loop-mediated isothermal amplification (LAMP), exhibit characteristics ideal for point-of-care (POC) applications, since their instrumentation is simpler in comparison with the standard method of polymerase chain reaction. Other key advantages of LAMP are robustness and the production of pyrophosphate in the presence of the target gene, enabling to detect the reaction products using the naked eye. Polymerase inhibitors, presented in clinical samples, do not affect the amplification process, making LAMP suitable for a simple sample-to-answer diagnostic systems with simplified sample preparation. In this review, we discuss the trends in miniaturized LAMP techniques, such as microfluidic, paper-based, and digital with their advantages and disadvantages, especially for POC applications alongside our opinion of the future development of miniaturized LAMP.

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Section: Temperature Cycling Amplificationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

LAMP-on-a-chip: Revising microfluidic platforms for loop-mediated DNA amplification

Zhang

Fohlerová

et al. 2019

TrAC Trends in Analytical Chemistry

181

126

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show abstract

“…Moreover, the design and development of conventional microfluidic devices are complex. Bulky external support systems such as pumps and tubing are required to deliver the samples into the microfluidic device, making many of them not suitable for practical field applications [10]. In this context, a suitable sample dispersion platform that has comparable performance to conventional PCR tubes and microfluidic devices but eliminating other disadvantages is of great interest.…”

Section: Introductionmentioning

confidence: 99%

Core-Shell Beads Made by Composite Liquid Marble Technology as A Versatile Microreactor for Polymerase Chain Reaction

et al. 2020

Self Cite

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Over the last three decades, the protocols and procedures of the DNA amplification technique, polymerase chain reaction (PCR), have been optimized and well developed. However, there have been no significant innovations in processes for sample dispersion for PCR that have reduced the amount of single-use or unrecyclable plastic waste produced. To address the issue of plastic waste, this paper reports the synthesis and successful use of a core-shell bead microreactor using photopolymerization of a composite liquid marble as a dispersion process. This platform uses the core-shell bead as a simple and effective sample dispersion medium that significantly reduces plastic waste generated compared to conventional PCR processes. Other improvements over conventional PCR processes of the novel dispersion platform include increasing the throughput capability, enhancing the performance and portability of the thermal cycler, and allowing for the contamination-free storage of samples after thermal cycling.

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“…These systems perform to a high level, but rely on multiple auxiliary devices leading to complex operating procedure and high-power usage. The future of dNAAT development; therefore, focuses on portability, integration, and miniaturization [12][13][14].In the course of NAAT, a series of heating schemes based on Peltier [15], near infrared (NIR) laser [16], acoustic waves [17], and other mechanisms were proposed and combined with fluorescence detection technology, thereby promoting the emergence of integrated qNAAT instruments including qPCR devices. Among these heating schemes, the NIR heating demonstrates the advantages of large heating area, fast speed, and easy integration with fluorescence detection unit.…”

mentioning

confidence: 99%

mentioning

confidence: 99%

An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA Amplification

Zhang

Zhao

et al. 2020

Micromachines

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The sensitive quantification of low-abundance nucleic acids holds importance for a range of clinical applications and biological studies. In this study, we describe a facile microfluidic chip for absolute DNA quantifications based on the digital loop-mediated isothermal amplification (digital LAMP) method. This microfluidic chip integrates a cross-flow channel for droplet generation with a micro-cavity for droplet tiling. DNA templates in the LAMP reagent were divided into ~20,000 water-in-oil droplets at the cross-flow channel. The droplets were then tiled in the micro-cavity for isothermal amplification and fluorescent detection. Different from the existing polydimethylsiloxane (PDMS) microfluidic chips, this study incorporates gold nanoparticles (AuNPs) into PDMS substrate through silica coating and dodecanol modification. The digital LAMP chip prepared by AuNPs-PDMS combines the benefits of the microstructure manufacturing performance of PDMS with the light-to-heat conversion advantages of AuNPs. Upon illumination with a near infrared (NIR) LED, the droplets were stably and efficiently heated by the AuNPs in PDMS. We further introduce an integrated device with a NIR heating unit and a fluorescent detection unit. The system could detect HBV (hepatitis B virus)-DNA at a concentration of 1 × 101 to 1 × 104 copies/μL. The LED-driven digital LAMP chip and the integrated device; therefore, demonstrate high accuracy and excellent performance for the absolute quantification of low-abundance nucleic acids, showing the advantages of integration, miniaturization, cost, and power consumption.

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Digital polymerase chain reaction technology – recent advances and future perspectives

Abstract: We critically evaluate the state of the art of the development of digital polymerase chain reaction systems.

Cited by 107 publications

References 149 publications

LAMP-on-a-chip: Revising microfluidic platforms for loop-mediated DNA amplification

LAMP-on-a-chip: Revising microfluidic platforms for loop-mediated DNA amplification

Core-Shell Beads Made by Composite Liquid Marble Technology as A Versatile Microreactor for Polymerase Chain Reaction

An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA Amplification

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