Dual detection system for cancer-associated point mutations assisted by a multiplexed LNA-based amperometric bioplatform coupled with rolling circle amplification

Sebuyoya, Ravery; Valverde, Alejandro; Moranova, Ludmila; Strmiskova, Johana; Hrstka, Roman; Montiel, Víctor Ruiz-Valdepeñas; Pingarrón, José M.; Barderas, Rodrigo; Campuzano, Susana; Bartosik, Martin

doi:10.1016/j.snb.2023.134375

Cited by 4 publications

(1 citation statement)

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“…For DNA amplification, isothermal techniques have recently gathered much attention from researchers owing to their rapidity, cost effectiveness, ease of use, and ability to be performed under constant temperature. Some common isothermal techniques include loop-mediated isothermal amplification (LAMP) [13][14][15], rolling circle amplification (RCA) [16,17], nucleic acid sequence-based amplification (NASBA) [18,19], and recombinase polymerase amplification (RPA) [20,21]. Among these listed isothermal techniques, LAMP serves as the most promising approach because it requires less enzyme compared to RPA.…”

Section: Introductionmentioning

confidence: 99%

Pipette-Free and Fully Integrated Paper Device Employing DNA Extraction, Isothermal Amplification, and Carmoisine-Based Colorimetric Detection for Determining Infectious Pathogens

Nguyen,

Lee

2023

Sensors

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A pipette-free and fully integrated device that can be used to accurately recognize the presence of infectious pathogens is an important and useful tool in point-of-care testing, particularly when aiming to decrease the unpredictable threats posed by disease outbreak. In this study, a paper device is developed to integrate the three main processes required for detecting infectious pathogens, including DNA extraction, loop-mediated isothermal amplification (LAMP), and detection. All key reagents, including sodium dodecyl sulfate (SDS), NaOH, LAMP reagents, and carmoisine, are placed on the paper device. The paper device is operated simply via sliding and folding without using any bulky equipment, and the results can be directly observed by the naked eye. The optimized concentrations of sodium dodecyl sulfate (SDS), sodium hydroxide (NaOH), and carmoisine were found to be 0.1%, 0.1 M, and 0.5 mg/mL, respectively. The paper device was used to detect Enterococcus faecium at concentrations as low as 102 CFU/mL within 60 min. Also, E. faecium spiked in milk was successfully detected using the paper device, demonstrating the feasible application in real sample analysis.

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