Label-free and Electrochemical Detection of Nucleic Acids Based on Isothermal Amplification in Combination with Solid-state pH Sensor

Tabata, Miyuki; Katayama, Yurika; Mannan, Fahmida; Seichi, Ayaka; Suzuki, Kôji; Goda, Tatsuro; Matsumoto, Akira; Miyahara, Yuji

doi:10.1016/j.proeng.2016.11.534

Cited by 9 publications

(6 citation statements)

References 5 publications

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“…In a nucleic acid extension reaction by polymerase, when one dNTP is bound to the template strand, one pyrophosphate and one proton are released as by-products. Needle-type Ir/IrOx electrode was prepared to monitor the pH change in PG-RCA [6]. To confirm pH sensitivity of the prepared micro pH sensor, three-point calibration was carried out with standard buffer solutions at pH 4.0, 7.0 and 9.2.…”

Section: Pg-rca-ph Sensingmentioning

confidence: 99%

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Label-free Nucleic Acid Amplification Detection using Electrochemical Sensors for Liquid Biopsy

Tabata

Miyahara

2019

J. Photopol. Sci. Technol.

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In recent years, isothermal nucleic acid amplification techniques have been developed as alternatives to polymerase chain reaction (PCR) requiring thermal cycles. The integration of isothermal amplification into electrical or electrochemical devices realizes high throughput nucleic acid-based assays and confers high sensitivity. In this research, we tried to develop electrical/electrochemical biosensors detecting nucleic acids aiming at application to future liquid biopsy. For detection of small DNAs or microRNAs (miRNAs), isothermal amplification methods were employed, and the signal changes based on the extension reaction by polymerases were detected using electrical/electrochemical devices, namely, a chronocoulometric sensor and a micro pH sensor. These small sensors based on electrochemical technique might be a promising tool in on-site detection of nucleic acids related to cancer diagnosis.

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Section: Pg-rca-ph Sensingmentioning

confidence: 99%

“…In this research, we propose the electrical/electrochemical biosensors in combination with isothermal nucleic acid amplification for quantitative evaluation of nucleic acids aiming at miniaturization of devices [5,6] Table 1.…”

Section: Introductionmentioning

confidence: 99%

Label-free Nucleic Acid Amplification Detection using Electrochemical Sensors for Liquid Biopsy

Tabata

Miyahara

2019

J. Photopol. Sci. Technol.

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“…While the metal oxide pH sensors have been used to detect pH of blood [8], skin [9], and urine [10], the sensors have not been explored as a reliable platform to detect H + from NA amplification. Recently, a label-free and electrochemical detection of NA based on primer-generation rolling circle amplification were developed on the iridium oxide pH sensor [11].…”

Section: Introductionmentioning

confidence: 99%

Identification of bacteria strains using the Recombinase Polymerase Amplification assay on a miniaturized solid-state pH sensor

Nguyen

Malhotra

Lau

et al. 2022

Preprint

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Rapid identification of bacteria based on nucleic acid amplification allows dealing with the detection of pathogens in clinical, food, and environmental samples. Amplification products must be detected and analyzed by external devices or integrated complicated optical systems. Here, we developed a solid-state pH electrode based on iridium oxide (IrO2) films to measure released hydrogen ions (H+) from isothermal nucleic acid (NA) amplification of bacterial samples. By recombinase polymerase amplification (RPA), we achieved rapid (< 15 min) and sensitive (<30 copies) detection with an accuracy of about 0.03 pH. The RPA-based hydrogen ion sensing assay shows higher specificity, sensitivity, and efficiency as the same polymerase chain reaction (PCR) methods. We initially used the RPA-based sensor to detect E. coli species in laboratory samples. Among, 27 random laboratory samples of E. coli samples, 6 were found to be DH5alpha, 9 BL21, 3 HB101, 6 TOP10, and 3 JM109. The electrical detection of amplification provides generally applicable techniques for the detection of nucleic acid amplification, enabling molecular diagnostic tests in the field and integrating data transmission to the mobile device. These results can be future developed into an efficient tool for rapid on-site detection of bacterial pathogens in clinical samples.

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“…The most widely studied all-solid-state ion-electrodes are comprised of conductive polymer materials, such as polypyrrole (PPy) [13], poly-3-octylthiophene (POT) [14], polyaniline (PANI) [15][16][17][18][19], poly(3,4-ethylenedioxythiophene) (PEDOT) [20,21], and so on. In addition, inorganic membrane electrodes are mostly insoluble salt electrodes [22], such as Ag/AgCl reference electrode [23], Ag/Ag 2 S electrode [24], and Ir/IrO x pH electrode [25]. Organic and inorganic composite membranes have been used to fabricate electrodes [26].…”

Section: Introductionmentioning

confidence: 99%

Doped PANI Coated Nano-Ag Electrode for Rapid In-Situ Detection of Bromide in Seawater

Wang

Zhou

et al. 2019

Coatings

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In this paper, we successfully fabricated a novel bromide ion selective electrode (Br-ISE), which was coated by bromine ion doped polyaniline as sensitive film. Using Ag wire as the substrate, a uniform and dense nano-silver layer was electroplated to enhance the specific surface area of the electrode. Subsequently, a polyaniline (PANI) film was coated onto the electrode by cyclic voltammetry in a 0.3 M aniline and 1 M HCl solution and was in-situ doped by 0.1 M KBr solution. The morphology and performance of the electrode were characterized by scanning electron microscopy (SEM), electrochemical impedance spectroscopy (EIS), and other electrochemical analysis methods, respectively. The prepared Br-ISE exhibited a wide linear dynamic range between 1.0 × 10−1 and 1.0 × 10−7 M with a near-Nernst slope of 57.33 mV/decade. In addition, the electrode possessed extremely fast response time (<1 s) and low impedance (300 Ω), high sensitivity, and good selectivity. The electrode potential drifted within 2 mV in 8 h. The lifespan was larger than three months.

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Label-free and Electrochemical Detection of Nucleic Acids Based on Isothermal Amplification in Combination with Solid-state pH Sensor

Cited by 9 publications

References 5 publications

Label-free Nucleic Acid Amplification Detection using Electrochemical Sensors for Liquid Biopsy

Label-free Nucleic Acid Amplification Detection using Electrochemical Sensors for Liquid Biopsy

Identification of bacteria strains using the Recombinase Polymerase Amplification assay on a miniaturized solid-state pH sensor

Doped PANI Coated Nano-Ag Electrode for Rapid In-Situ Detection of Bromide in Seawater

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