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

Solid-state electrochemical sensors are developing as a new platform for liquid biopsy, combining detection and analysis of nucleic acids with isothermal nucleic acid amplification reactions.

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Low-cost colorimetric reader and label-free strategy for user-friendly detection of nucleic acid amplification products

Sensors and Actuators B Chemical ◽

10.1016/j.snb.2021.130523 ◽

2021 ◽

pp. 130523

Author(s):

Wenzhi Tang ◽

Meng Zhang ◽

Tianli Yue ◽

Xin Wang ◽

Zhonghong Li

Keyword(s):

Nucleic Acid ◽

Low Cost ◽

Nucleic Acid Amplification ◽

Label Free ◽

User Friendly

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Development of a Portable SPR Sensor for Nucleic Acid Detection

Micromachines ◽

10.3390/mi11050526 ◽

2020 ◽

Vol 11 (5) ◽

pp. 526 ◽

Cited By ~ 1

Author(s):

Yafeng Huang ◽

Lulu Zhang ◽

Hao Zhang ◽

Yichen Li ◽

Luyao Liu ◽

...

Keyword(s):

Nucleic Acid ◽

Incident Angle ◽

Incident Light ◽

High Sensitivity ◽

Nucleic Acid Amplification ◽

Nucleic Acid Hybridization ◽

Nucleic Acid Detection ◽

Scanning System ◽

Label Free ◽

Spr Sensor

Nucleic acid detection is of great significance in clinical diagnosis, environmental monitoring and food safety. Compared with the traditional nucleic acid amplification detection method, surface plasmon resonance (SPR) sensing technology has the advantages of being label-free, having simple operation, and providing real-time detection. However, the angle scanning system in many SPR angle modulation detection applications usually requires a high-resolution stepper motor and complex mechanical structure to adjust the angle. In this paper, a portable multi-angle scanning SPR sensor was designed. The sensor only uses one stepping motor to rotate a belt, and the belt pulls the mechanical linkages of incident light and reflected light to move in opposite directions for achieving the SPR angle scanning mode that keeps the incident angle and reflected angle equal. The sensor has an angle scanning accuracy of 0.002°, response sensitivity of 3.72 × 10−6 RIU (refractive index unit), and an angle scanning range of 30°–74°. The overall size of the system is only 480 mm × 150 mm × 180 mm. The portable SPR sensor was used to detect nucleic acid hybridization on a gold film chip modified with bovine serum albumin (BSA). The result revealed that the sensor had high sensitivity and fast response, and could successfully accomplish the hybridization detection of target DNA solution of 0.01 μmol/mL.

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1029. Torus Synestia Nucleic Acid Analysis Platform for Fast, High Multiplex Analysis of Nucleic Acids With Single-Nucleotide Discrimination Level

Open Forum Infectious Diseases ◽

10.1093/ofid/ofab466.1223 ◽

2021 ◽

Vol 8 (Supplement_1) ◽

pp. S605-S605

Author(s):

Tyler Rockwood ◽

Andrew Sullivan ◽

Jahnavi Gandhi ◽

Sarah Gruszka ◽

Brian Turczyk ◽

...

Keyword(s):

Nucleic Acid ◽

Target Detection ◽

Bacterial Species ◽

Nucleic Acid Amplification ◽

Detection Sensitivity ◽

Label Free ◽

Single Nucleotide ◽

Advantages And Disadvantages ◽

Tract Infections ◽

Dna 2

Abstract Background Nucleic acid amplification testing (NAAT) is an essential tool both for biomedical research and for clinical molecular diagnostics. Currently, there are multiple NAAT platforms available, each offering certain performance and utility advantages and disadvantages as compared to each other. Next generation NAAT platforms aim to deliver increased target detection sensitivity and specificity, low limits of target detection, quantitative high multiplex target capacity, rapid time to results, and simple sample-to-answer workflow. Methods Here we describe the Torus Synestia System, a NAAT platform capable of rapid, highly multiplexed amplification and detection of both DNA and RNA targets. The platform comprises a small, portable (~ 2kg) amplification and detection device and a disposable single-use cartridge housing a PCR amplification chamber with an integrated label-free microarray for real-time data acquisition and interpretation. The platform offers a 30-min turnaround time with a detection limit of 10 DNA/RNA molecules per assay and single nucleotide discrimination. Results We demonstrate the Synestia System performance and utility with three distinct molecular applications: 1) detection of 20 genetic loci and 30 single nucleotide polymorphisms in human genomic DNA; 2) detection and genotyping of 43 unique bacterial species associated with human urinary tract infections; and 3) detection and profiling human respiratory viral pathogens including SARS-CoV-1/2, seasonal coronaviruses, Influenza A/B, and human respiratory syncytial viruses. In addition, the single-nucleotide specificity of our label-free microarray probes allowed for robust identification and discrimination of newly emerging SARS-CoV-2 lineages, such as B.1.1.7 (a.k.a. UK), B.1.351 (a.k.a. South African), P.1 (a.k.a. Brazilian), and B.1.617 (a.k.a. Indian). Conclusion The Torus Synestia System has broad applicability in both clinical and research environments. We are confident that the Torus Synestia System will revolutionize syndromic diagnostics at the point of care (PoC) and lead to improved response times during future epidemic and pandemic pathogen outbreaks. Disclosures All Authors: No reported disclosures

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A rapid and label-free DNA-based interference reduction nucleic acid amplification strategy for viral RNA detection

Biosensors and Bioelectronics ◽

10.1016/j.bios.2021.113829 ◽

2021 ◽

pp. 113829

Author(s):

Feng Chen ◽

Guodong Li ◽

Chun Wu ◽

Wanhe Wang ◽

Dik-Lung Ma ◽

...

Keyword(s):

Nucleic Acid ◽

Viral Rna ◽

Nucleic Acid Amplification ◽

Label Free ◽

Rna Detection ◽

Interference Reduction ◽

Free Dna ◽

Amplification Strategy

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A Label-Free Impedimetric Genosensor for the Nucleic Acid Amplification-Free Detection of Extracted RNA of Dengue Virus

Sensors ◽

10.3390/s20133728 ◽

2020 ◽

Vol 20 (13) ◽

pp. 3728

Author(s):

Ching-Chou Wu ◽

Hao-Yu Yen ◽

Lu-Ting Lai ◽

Guey-Chuen Perng ◽

Cheng-Rei Lee ◽

...

Keyword(s):

Nucleic Acid ◽

Dengue Virus ◽

Electrochemical Impedance ◽

Denv Infection ◽

Viral Disease ◽

Diagnostic Methods ◽

Nucleic Acid Amplification ◽

Great Promise ◽

Label Free ◽

Hybridization Efficiency

Developing rapid and sensitive diagnostic methods for dengue virus (DENV) infection is of prime priority because DENV infection is the most prevalent mosquito-borne viral disease. This work proposes an electrochemical impedance spectroscopy (EIS)-based genosensor for the label-free and nucleic acid amplification-free detection of extracted DENV RNA intended for a sensitive diagnosis of DENV infection. A concentration ratio of 0.04 mM 6-mercaptohexanoic acid (MHA) to 1 mM 6-mercapto-1-hexanol (MCH) was selected to modify thin-film gold electrodes as a link to control the coverage of self-designed probe DNA (pDNA) at a density of 4.5 ± 0.4 × 1011 pDNA/cm2. The pDNA/MHA/MCH-modified genosensors are proven to improve the hybridization efficiency of a synthetic 160-mer target DNA (160mtDNA) with a 140-mer electrode side overhang as compared to other MHA/MCH ratio-modified genosensors. The MHA(0.04 mM)/MCH(1 mM)-modified genosensors also present good hybridization efficiency with the extracted DENV serotype 1 (DENV1) RNA samples, having the same electrode side overhangs with the 160mtDNA, showing a low detection limit of 20 plaque forming units (PFU)/mL, a linear range of 102–105 PFU/mL and good selectivity for DENV1. The pDNA density-controlled method has great promise to construct sensitive genosensors based on the hybridization of extracted DENV nucleic acids.

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