Duplex-specific nuclease-mediated target recycling amplification for fluorescence detection of microRNA

2019 ◽  
Vol 11 (2) ◽  
pp. 200-204 ◽  
Author(s):  
Lin Tan ◽  
Liu Xu ◽  
Jin-Wen Liu ◽  
Li-Juan Tang ◽  
Hao Tang ◽  
...  

Isothermal amplification techniques for nucleic acid detection have drawn increasing interest recently due to the simplicity and low-cost of instruments.

RSC Advances ◽  
2016 ◽  
Vol 6 (13) ◽  
pp. 10650-10654 ◽  
Author(s):  
Haiping Wei ◽  
Yan Wang ◽  
Hong Zhang ◽  
Haiyan Zhao ◽  
Wei Jiang

A sensitive strategy based on Au nanoparticles fluorescence switch-mediated target recycling amplification for nucleic acid detection.


2013 ◽  
Vol 59 (2) ◽  
pp. 436-439 ◽  
Author(s):  
Martin Jensen Søe ◽  
Mikkel Rohde ◽  
Jens Mikkelsen ◽  
Peter Warthoe

BACKGROUND Nucleic acid tests that can simultaneously detect multiple targets with high sensitivity, specificity, and speed are highly desirable. To meet this need, we developed a new approach we call the isoPCR method. METHODS The isoPCR method is a 2-stage nested-like nucleic acid amplification method that combines a single multiplex preamplification PCR with subsequent distinct detection of specific targets by use of isothermal amplification. We compared isoPCR to nested quantitative PCR (qPCR), loop-mediated isothermal amplification (LAMP), and nested LAMP (PCR followed by LAMP), for detection of DNA from Candida glabrata. We evaluated the method's multiplex capability for detecting low copy numbers of pathogens commonly involved in sepsis. RESULTS IsoPCR provided detection of 1 copy of Candida glabrata, an LOD that was 5-fold lower than a nested qPCR assay (5 copies), while the amplification time was simultaneously halved. Similarly, the LOD for isoPCR was lower than that for a LAMP assay (1000 copies) and a nested LAMP assay (5 copies). IsoPCR required recognition of 6 regions for detection, thereby providing a theoretically higher specificity compared to nested qPCR (4 regions). The isoPCR multiplexing capability was demonstrated by simultaneous detection of 4 pathogens with individual LODs of 10 copies or fewer. Furthermore, the specificity of isoPCR was demonstrated by successful pathogen detection from samples with more than 1 pathogen present. CONCLUSIONS IsoPCR provides a molecular diagnostic tool for multiplex nucleic acid detection, with an LOD down to 1 copy, high theoretical specificity, and halving of the amplification time compared to a nested qPCR assay.


PLoS ONE ◽  
2019 ◽  
Vol 14 (12) ◽  
pp. e0220091 ◽  
Author(s):  
Florian Katzmeier ◽  
Lukas Aufinger ◽  
Aurore Dupin ◽  
Jorge Quintero ◽  
Matthias Lenz ◽  
...  

2016 ◽  
Vol 52 (74) ◽  
pp. 11108-11111 ◽  
Author(s):  
Xinya Sun ◽  
Li Wang ◽  
Mingsha Zhao ◽  
Changzhi Zhao ◽  
Shufeng Liu

An autonomous target recycling and cascade circular exponential amplification strategy was proposed for the one-pot, isothermal and ultrasensitive detection of target DNA.


Lab on a Chip ◽  
2015 ◽  
Vol 15 (7) ◽  
pp. 1697-1707 ◽  
Author(s):  
Mark D. Borysiak ◽  
Kevin W. Kimura ◽  
Jonathan D. Posner

The NAIL device integrates isotachophoresis and loop-mediated isothermal amplification (LAMP) with mobile phone detection to extract, amplify, and detect nucleic acids from complex matrices in less than one hour.


2014 ◽  
Vol 43 (17) ◽  
pp. 6405-6438 ◽  
Author(s):  
Yulia V. Gerasimova ◽  
Dmitry M. Kolpashchikov

Enzyme-assisted target recycling (EATR) is a signal amplification technique that can find application in PCR-free nucleic acid detection.


2020 ◽  
Author(s):  
Eduardo Nogueira Cunha ◽  
Maria Fernanda Bezerra de Souza ◽  
Daniel Carlos Ferreira Lanza ◽  
João Paulo Matos Santos Lima

ABSTRACTNucleic acid detection by electrophoresis is still a quick and accessible technique for many diagnosis methods, primarily at research laboratories or at the point of care units. Standard protocols detect DNA/RNA molecules through specific bound chemical dyes using a UV-transilluminator or UV-photo documentation system. However, the acquisition costs and availability of these devices, mainly the ones with photography and internet connection capabilities, can be prohibitive, especially in developing countries public health units. Also, ultraviolet radiation is a common additional risk factor to professionals that use electrophoresis-based nucleic acid detection. With that in mind, this work describes the development of a low-cost DNA/RNA detection smart system capable of obtaining qualitative and semi-quantitative data from gel analysis. The proposed device explores the visible light absorption range of commonly used DNA/RNA dyes using readily available parts, and simple manufacturing processes, such as light-emitting diodes (LEDs) and 3D impression. By applying IoT techniques, our system covers a wide range of color spectrum in order to detect bands from various commercially used dyes, using Bluetooth communication and a smartphone for hardware control, image capturing, and sharing. The project also enables process scalability and has low manufacturing and maintenance costs. The use of LEDs at the visible spectrum can achieve very reproducible images, providing a high potential for rapid and point-of-care diagnostics as well as applications in several fields such as healthcare, agriculture, and aquaculture.


2016 ◽  
Vol 2 (3) ◽  
pp. 278-294 ◽  
Author(s):  
Mohammadali Safavieh ◽  
Manoj K. Kanakasabapathy ◽  
Farhang Tarlan ◽  
Minhaz U. Ahmed ◽  
Mohammed Zourob ◽  
...  

2006 ◽  
Vol 315-316 ◽  
pp. 469-473
Author(s):  
Ji Jun Zhu ◽  
H.N. Shi ◽  
J. Cheng ◽  
X.Y. Wei ◽  
Zu Hong Lu

This paper introduces a kind of new homemade and low cost multi-channel nucleic acid pyrosequence detector for at least ninety-six channels and presents the detail of related software and hardware development. We construct a kind of automatic instrument to fulfill the pyrosequencing processes. First we select the X-86 personal computer as host computer, the AT89C51 micro-controller as slave computer, the PMT (photoelectric multiply tube) as photoelectric transformation equipment, and the HY-6022 as data sampling device; Second we use the Visual C++ 6.0 as coding tools to design the measure and control system based on Windows 2000 operating system; Third we sample the fluorescent signal in all of the cuvettes during the reaction between nucleic acid and reagent; Last we analyze these data to realize the function of the multi-channel nucleic acid detection. In this paper the whole instrument design and key parts design are both introduced such as the liquid injection process and related structure design, the communication module between the host personal computer and the MCS51, the high sensitivity multi-channel detector (at least 96 channels, the sensitivity is 2.45×10-9w) etc. The result of the instrument for two channels data processing is also reported in this paper.


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