Biosensors based on functional nucleic acids and isothermal amplification techniques

Cao, Xiuen; Chen, Chuanpin; Zhu, Qubo

doi:10.1016/j.talanta.2022.123977

Cited by 37 publications

(18 citation statements)

References 186 publications

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“…The selected gene targets for LAMP and RPA have different lengths and GC content. However, the mechanism of two amplifications is so different that even within the same gene it is impossible for RPA and LAMP to obtain the same amplicons [ 49 ]; therefore, given the same copy number and proven efficiency of primers, different gene localization is not a significant factor.…”

Section: Resultsmentioning

confidence: 99%

Comparison of Biosensing Methods Based on Different Isothermal Amplification Strategies: A Case Study with Erwinia amylovora

et al. 2022

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Isothermal amplifications allow for the highly sensitive detection of nucleic acids, bypassing the use of instrumental thermal cycling. This work aimed to carry out an experimental comparison of the four most promising techniques: recombinase polymerase amplification (RPA) and loop-mediated isothermal amplification (LAMP) coupled with lateral flow test or coupled with additional amplification based on CRISPR/Cas12a resulting from the fluorescence of the Cas12a-cleaved probe. To compare the four amplification techniques, we chose the bacterial phytopathogen Erwinia amylovora (causative agent of fire blight), which has a quarantine significance in many countries and possesses a serious threat to agriculture. Three genes were chosen as the targets and primers were selected for each one (two for RPA and six for LAMP). They were functionalized by labels (biotin, fluorescein) at the 5′ ends for amplicons recognition by LFT. As a result, we developed LAMP-LFT, LAMP-CRISPR/Cas, RPA-LFT, and RPA-CRISPR/Cas for E. amylovora detection. The detection limit was 104 CFU/mL for LAMP-LFT, 103 CFU/mL for LAMP-CRISPR/Cas, and 102 CFU/mL for RPA-LFT and RPA-CRISPR/Cas. The results of four developed test systems were verified by qPCR on a panel of real samples. The developed assays based on RPA, LAMP, CRISPR/Cas12a, and LFT are rapid (30–55 min), user-friendly, and highly sensitive for E. amylovora detection. All proposed detection methods can be applied to fire blight diagnosis and effective management of this disease.

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Section: Resultsmentioning

confidence: 99%

Comparison of Biosensing Methods Based on Different Isothermal Amplification Strategies: A Case Study with Erwinia amylovora

et al. 2022

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“…Compared to existing food safety detection methods, biosensors offer significant advantages such as low cost, rapid analysis, and ease of use [ 33 , 34 ]. Due to the advantages of biocompatibility, programmability and good external response, stimulus-responsive DNA hydrogel can be used as a recognition element for biosensors to improve the sensitivity and specificity [ 35 ]. In this review, we will first outline general strategies for constructing stimulus-responsive DNA hydrogels.…”

Section: Introductionmentioning

confidence: 99%

Stimulus-Responsive DNA Hydrogel Biosensors for Food Safety Detection

Wang

Lai

et al. 2023

Biosensors

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Food safety has always been a major global challenge to human health and the effective detection of harmful substances in food can reduce the risk to human health. However, the food industry has been plagued by a lack of effective and sensitive safety monitoring methods due to the tension between the cost and effectiveness of monitoring. DNA-based hydrogels combine the advantages of biocompatibility, programmability, the molecular recognition of DNA molecules, and the hydrophilicity of hydrogels, making them a hotspot in the research field of new nanomaterials. The stimulus response property greatly broadens the function and application range of DNA hydrogel. In recent years, DNA hydrogels based on stimulus-responsive mechanisms have been widely applied in the field of biosensing for the detection of a variety of target substances, including various food contaminants. In this review, we describe the recent advances in the preparation of stimuli-responsive DNA hydrogels, highlighting the progress of its application in food safety detection. Finally, we also discuss the challenges and future application of stimulus-responsive DNA hydrogels.

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Section: ■ Introductionmentioning

confidence: 99%

“…Isothermal amplification combined with DNAzyme walker has the potential to mitigate the challenge due to the ease in integration with additional components and high amplification efficiency under a constant temperature. , In particular, rolling circle amplification (RCA) is one of the most attractive techniques for universal low-abundance molecular detection benefiting from its amplified products containing thousands of periodic repeat sequences. − For example, Ju et al presented a cascade DNA reaction by hybridizing RCA-produced nanowire with hairpin probes, which could be used for the detection of different mRNAs . The Xu and Huang group developed a dual-mode RCA biosensing platform, achieving electrochemical and colorimetric detection of microRNA with detection limits of 0.15 and 33 fM .…”

Section: Introductionmentioning

confidence: 99%

Universal Detection and Imaging for Multiple Targets by Coupling Target-Primed Nicking-Enhanced Rolling Circle Amplification with Self-Powered DNAzyme Walker

Zhang

et al. 2023

Anal. Chem.

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Although promising in monitoring low-abundance analytes, most of the DNAzyme walker is only responsive to a specific target. Herein, a universal, ready-to-use platform is developed by coupling nicking-enhanced rolling circle amplification and a self-powered DNAzyme walker (NERSD). It addressed the issues that DNAzyme strands need to be specifically designed for different biosensing system, allowing highly sensitive analysis of various targets with the same DNAzyme walker components. It is also specific owing to target-dependent ligation of the padlock probe and precise cleavage of a substrate by a DNAzyme strand. As typically demonstrated, the strategy has an equivalent capacity with the qRT-PCR kit in distinguishing plasma miR-21 levels of breast cancer patients from normal subjects and is able to differentiate intracellular miR-21 and ATP levels by confocal imaging. The approach characteristic of programmability, flexibility, and generality indicated the potential in all kinds of biosensing and imaging platform.

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Biosensors based on functional nucleic acids and isothermal amplification techniques

Cited by 37 publications

References 186 publications

Comparison of Biosensing Methods Based on Different Isothermal Amplification Strategies: A Case Study with Erwinia amylovora

Comparison of Biosensing Methods Based on Different Isothermal Amplification Strategies: A Case Study with Erwinia amylovora

Stimulus-Responsive DNA Hydrogel Biosensors for Food Safety Detection

Universal Detection and Imaging for Multiple Targets by Coupling Target-Primed Nicking-Enhanced Rolling Circle Amplification with Self-Powered DNAzyme Walker

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