Closed dumbbell mediated isothermal amplification of nucleic acids for DNA diagnostic assays

Talanta ◽  
2022 ◽  
pp. 123217
Author(s):  
Rui Mao ◽  
Tianzuo Wang ◽  
Yue Zhao ◽  
Xinyao Wu ◽  
Shun Zhang ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Isothermal Amplification ◽  
Diagnostic Assays

scholarly journals An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA Amplification

Micromachines ◽  
2020 ◽  
Vol 11 (2) ◽  
pp. 177 ◽  
Author(s):  
Zengming Zhang ◽  
Shuhao Zhao ◽  
Fei Hu ◽  
Guangpu Yang ◽  
Juan Li ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Microfluidic Chip ◽  
Cross Flow ◽  
Flow Channel ◽  
Isothermal Amplification ◽  
Fluorescent Detection ◽  
Lamp Method ◽  
Biological Studies ◽  
Micro Cavity ◽  
Integrated Device

The sensitive quantification of low-abundance nucleic acids holds importance for a range of clinical applications and biological studies. In this study, we describe a facile microfluidic chip for absolute DNA quantifications based on the digital loop-mediated isothermal amplification (digital LAMP) method. This microfluidic chip integrates a cross-flow channel for droplet generation with a micro-cavity for droplet tiling. DNA templates in the LAMP reagent were divided into ~20,000 water-in-oil droplets at the cross-flow channel. The droplets were then tiled in the micro-cavity for isothermal amplification and fluorescent detection. Different from the existing polydimethylsiloxane (PDMS) microfluidic chips, this study incorporates gold nanoparticles (AuNPs) into PDMS substrate through silica coating and dodecanol modification. The digital LAMP chip prepared by AuNPs-PDMS combines the benefits of the microstructure manufacturing performance of PDMS with the light-to-heat conversion advantages of AuNPs. Upon illumination with a near infrared (NIR) LED, the droplets were stably and efficiently heated by the AuNPs in PDMS. We further introduce an integrated device with a NIR heating unit and a fluorescent detection unit. The system could detect HBV (hepatitis B virus)-DNA at a concentration of 1 × 101 to 1 × 104 copies/μL. The LED-driven digital LAMP chip and the integrated device; therefore, demonstrate high accuracy and excellent performance for the absolute quantification of low-abundance nucleic acids, showing the advantages of integration, miniaturization, cost, and power consumption.

scholarly journals A Sensitive Potentiometric Sensor for Isothermal Amplification-Coupled Detection of Nucleic Acids

Sensors ◽  
2018 ◽  
Vol 18 (7) ◽  
pp. 2277
Author(s):  
Kang-Ho Lee ◽  
Dongkyu Lee ◽  
Jongsu Yoon ◽  
Ohwon Kwon ◽  
Jaejong Lee
Keyword(s):  
Nucleic Acids ◽  
Real Time ◽  
Surface Potential ◽  
Pathogenic Bacteria ◽  
Field Effect Transistors ◽  
Potentiometric Sensor ◽  
Isothermal Amplification ◽  
Ring Oscillator ◽  
Oxide Semiconductor ◽  
High Signal

A disposable potentiometric sensor was newly developed for the amplification-coupled detection of nucleic acids. The hydrogen-ion is generally released during isothermal amplification of nucleic acids. The surface potential on the oxide-functionalized electrode of the extended gate was directly measured using full electrical circuits with the commercial metal-oxide semiconductor field-effect transistors (MOSFETs) and ring oscillator components, which resulted in cost-effective, portable and scalable real-time nucleic acid analysis. The current-starved ring oscillator changes surface potential to its frequency depending on the square of the variation in pH with a high signal-to-noise ratio during isothermal amplification. The device achieves a conversion rate of 20.5 kHz/mV and a detection resolution of 200 µV for the surface potential. It is demonstrated that the sensor successfully monitors in real-time isothermal amplification of the extracted nucleic acids from Salmonella pathogenic bacteria. The in situ variations in the frequency of the pH-sensitive sensor were compared with the results of both a conventional optical device and pH-meter during isothermal amplification.

Competitive annealing mediated isothermal amplification of nucleic acids

The Analyst ◽  
2018 ◽  
Vol 143 (3) ◽  
pp. 639-642 ◽  
Author(s):  
Rui Mao ◽  
Lifei Qi ◽  
Jianjun Li ◽  
Ming Sun ◽  
Zhuo Wang ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
High Specificity ◽  
Isothermal Amplification ◽  
Amplification Method

A novel nucleic acid isothermal amplification method with high specificity, efficiency and rapidity was developed.

scholarly journals Critical review of methods for isothermal amplification of nucleic acids for environmental analysis

2020 ◽  
Vol 179 ◽  
pp. 106099
Author(s):  
Dana M. Nieuwkerk ◽  
Asja Korajkic ◽  
Erika L. Valdespino ◽  
Michael P. Herrmann ◽  
Valerie J. Harwood
Keyword(s):  
Nucleic Acids ◽  
Environmental Analysis ◽  
Critical Review ◽  
Isothermal Amplification

Point-of-Care Multiplexed Assays of Nucleic Acids Using Microcapillary-based Loop-Mediated Isothermal Amplification

2014 ◽  
Vol 86 (14) ◽  
pp. 7057-7062 ◽  
Author(s):  
Yi Zhang ◽  
Lu Zhang ◽  
Jiashu Sun ◽  
Yulei Liu ◽  
Xingjie Ma ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Point Of Care ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification

scholarly journals Enhancing colorimetric loop-mediated isothermal amplification speed and sensitivity with guanidine chloride

BioTechniques ◽  
2020 ◽  
Vol 69 (3) ◽  
pp. 178-185 ◽  
Author(s):  
Yinhua Zhang ◽  
Guoping Ren ◽  
Jackson Buss ◽  
Andrew J Barry ◽  
Gregory C Patton ◽  
...  
Keyword(s):  
Guanidine Hydrochloride ◽  
Isothermal Amplification ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Simple Method ◽  
Diagnostic Assays ◽  
Loop Mediated Isothermal Amplification ◽  
Dna And Rna ◽  
Versatile Technique ◽  
Primer Sets

Loop-mediated isothermal amplification (LAMP) is a versatile technique for detection of target DNA and RNA, enabling rapid molecular diagnostic assays with minimal equipment. The global SARS-CoV-2 pandemic has presented an urgent need for new and better diagnostic methods, with colorimetric LAMP utilized in numerous studies for SARS-CoV-2 detection. However, the sensitivity of colorimetric LAMP in early reports has been below that of the standard RT-qPCR tests, and we sought to improve performance. Here we report the use of guanidine hydrochloride and combined primer sets to increase speed and sensitivity in colorimetric LAMP, bringing this simple method up to the standards of sophisticated techniques and enabling accurate, high-throughput diagnostics.

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