Deskilled and Rapid Drug-Resistant Gene Detection by Centrifugal Force-Assisted Thermal Convection PCR Device

Espulgar, Wilfred; Saito, Masato; Takahashi, Kazuya; Ushiro, Sakiko; Yamamoto, Naoki; Akeda, Yukihiro; Hamaguchi, Shigeto; Tomono, Kazunori; Tamiya, Eiichi

doi:10.3390/s21041225

Cited by 6 publications

(5 citation statements)

References 30 publications

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“…However, the right meniscus was not moving in parallel with the other meniscus due to fabrication problems. Aside from the fabrication issue, the microchannel behaved as was expected according to previous work (13,20).…”

Section: Thermal Convection Flow On CDsupporting

confidence: 67%

(Digital Presentation) An Electrochemical Sensor in a Disc for Real-Time PCR Assisted with Thermal Convective Flow

Romero-Soto¹,

Aeinehvand²,

Madou³

et al. 2022

ECS Trans.

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Multiple microfluidic devices incorporating real-time polymerase chain reaction (PCR) for the quantification of DNA targets have been reported. They show features such as fast analysis time, reduced reagent consumption, and portability. Integration of real-time PCR in a centrifugal microfluidic disc has been also proposed due to the ease of automation and parallelization of microfluidic operations. Previously, it has been demonstrated the implementation of thermal convective PCR (TC-PCR) in a CD using a ring-structured microchannel. In this case, the microchannel is supplied with the temperature necessary to perform the complete thermal cycle: Denaturation, annealing, and extension. The flow generated by thermal convection, in combination with the centrifugal force moves the sample through the entire microchannel due to the difference in densities. In this work, we present the implementation in a CD of an electrochemical sensor combined with TC-PCR to quantify the amplification yield in real-time. We present advances related to the calibration of the electrochemical sensor, and the implementation of the TC-PCR in a CD as well as a preliminary simulation in COMSOL.

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Section: Thermal Convection Flow On CDsupporting

confidence: 67%

(Digital Presentation) An Electrochemical Sensor in a Disc for Real-Time PCR Assisted with Thermal Convective Flow

Romero-Soto¹,

Aeinehvand²,

Madou³

et al. 2022

ECS Trans.

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“… Lok et al (2012a) reported on the design of a magnetically actuated circular closed-loop PCR chip, in which a ferrofluid plug is driven by an external magnet that moves along the microchannel and four loops can carry out PCRs simultaneously. Espulgar et al (2021) fabricated an identical closed-loop channel on a centrifugal disk-like chip; this design can apply 106 thermal cycles in 15 min. The drawbacks of closed-loop PCR chips are the inconvenience of retrieving the solution and an asymmetrical reaction procedure in concentric channels when high-throughput testing is attempted.…”

Section: Rapid Dna Amplification Methodsmentioning

confidence: 99%

“…In many studies, fluorescent dye (such as calcein or SYBR Green) has been added to create a more conspicuous light green color for easy detection. Zhang M. et al (2021) embedded calcein-soaked paper in the reaction chamber for direct detection of LAMP products.…”

Section: Fluorescent and Colorimetric Detectionmentioning

confidence: 99%

Rapid on-site nucleic acid testing: On-chip sample preparation, amplification, and detection, and their integration into all-in-one systems

Wang

Jiang

Pan

et al. 2023

Front. Bioeng. Biotechnol.

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As nucleic acid testing is playing a vital role in increasingly many research fields, the need for rapid on-site testing methods is also increasing. The test procedure often consists of three steps: Sample preparation, amplification, and detection. This review covers recent advances in on-chip methods for each of these three steps and explains the principles underlying related methods. The sample preparation process is further divided into cell lysis and nucleic acid purification, and methods for the integration of these two steps on a single chip are discussed. Under amplification, on-chip studies based on PCR and isothermal amplification are covered. Three isothermal amplification methods reported to have good resistance to PCR inhibitors are selected for discussion due to their potential for use in direct amplification. Chip designs and novel strategies employed to achieve rapid extraction/amplification with satisfactory efficiency are discussed. Four detection methods providing rapid responses (fluorescent, optical, and electrochemical detection methods, plus lateral flow assay) are evaluated for their potential in rapid on-site detection. In the final section, we discuss strategies to improve the speed of the entire procedure and to integrate all three steps onto a single chip; we also comment on recent advances, and on obstacles to reducing the cost of chip manufacture and achieving mass production. We conclude that future trends will focus on effective nucleic acid extraction via combined methods and direct amplification via isothermal methods.

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“…For a microfluidic PCR, the thermal cycling time is calculated in minutes, and the size of the reactor is required in μL, whereas a microfluidic PCR is defined as γ [ 21 ]. The first type of reactor is termed as a stationary reactor, forced continuous flow PCR is the second type of reactor and the third type of reactor is composed of a free heat deportation system [ 21 , 22 , 23 , 24 ].…”

Section: Microfluidic Technology and Poctsmentioning

confidence: 99%

Prospects of Microfluidic Technology in Nucleic Acid Detection Approaches

et al. 2023

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Conventional diagnostic techniques are based on the utilization of analyte sampling, sensing and signaling on separate platforms for detection purposes, which must be integrated to a single step procedure in point of care (POC) testing devices. Due to the expeditious nature of microfluidic platforms, the trend has been shifted toward the implementation of these systems for the detection of analytes in biochemical, clinical and food technology. Microfluidic systems molded with substances such as polymers or glass offer the specific and sensitive detection of infectious and noninfectious diseases by providing innumerable benefits, including less cost, good biological affinity, strong capillary action and simple process of fabrication. In the case of nanosensors for nucleic acid detection, some challenges need to be addressed, such as cellular lysis, isolation and amplification of nucleic acid before its detection. To avoid the utilization of laborious steps for executing these processes, advances have been deployed in this perspective for on-chip sample preparation, amplification and detection by the introduction of an emerging field of modular microfluidics that has multiple advantages over integrated microfluidics. This review emphasizes the significance of microfluidic technology for the nucleic acid detection of infectious and non-infectious diseases. The implementation of isothermal amplification in conjunction with the lateral flow assay greatly increases the binding efficiency of nanoparticles and biomolecules and improves the limit of detection and sensitivity. Most importantly, the deployment of paper-based material made of cellulose reduces the overall cost. Microfluidic technology in nucleic acid testing has been discussed by explicating its applications in different fields. Next-generation diagnostic methods can be improved by using CRISPR/Cas technology in microfluidic systems. This review concludes with the comparison and future prospects of various microfluidic systems, detection methods and plasma separation techniques used in microfluidic devices.

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Deskilled and Rapid Drug-Resistant Gene Detection by Centrifugal Force-Assisted Thermal Convection PCR Device

Cited by 6 publications

References 30 publications

(Digital Presentation) An Electrochemical Sensor in a Disc for Real-Time PCR Assisted with Thermal Convective Flow

(Digital Presentation) An Electrochemical Sensor in a Disc for Real-Time PCR Assisted with Thermal Convective Flow

Rapid on-site nucleic acid testing: On-chip sample preparation, amplification, and detection, and their integration into all-in-one systems

Prospects of Microfluidic Technology in Nucleic Acid Detection Approaches

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