Miniaturized isothermal nucleic acid amplification, a review

Abstract

Micro-Total Analysis Systems (µTAS) for use in on-site rapid detection of DNA or RNA are increasingly being developed. Here, amplification of the target sequence is key to increasing sensitivity, enabling single-cell and few-copy nucleic acid detection. The several advantages to miniaturizing amplification reactions and coupling them with sample preparation and detection on the same chip are well known and include fewer manual steps, preventing contamination, and significantly reducing the volume of expensive reagents. To-date, the majority of miniaturized systems for nucleic acid analysis have used the polymerase chain reaction (PCR) for amplification and those systems are covered in previous reviews. This review provides a thorough overview of miniaturized analysis systems using alternatives to PCR, specifically isothermal amplification reactions. With no need for thermal cycling, isothermal microsystems can be designed to be simple and low-energy consuming and therefore may outperform PCR in portable, battery-operated detection systems in the future. The main isothermal methods as miniaturized systems reviewed here include nucleic acid sequence-based amplification (NASBA), loop-mediated isothermal amplification (LAMP), helicase-dependent amplification (HDA), rolling circle amplification (RCA), and strand displacement amplification (SDA). Also, important design criteria for the miniaturized devices are discussed. Finally, the potential of miniaturization of some new isothermal methods such as the exponential amplification reaction (EXPAR), isothermal and chimeric primer-initiated amplification of nucleic acids (ICANs), signal-mediated amplification of RNA technology (SMART) and others is presented.

Loop-Mediated Isothermal Amplification (LAMP) Assay for GMO on: Recent Progresses and Future Perspectives

“…It is less sensitive to inhibitors [18] [19], no need of complicate equipment, simple to operate, visual result, and easy to be developed into commercially available detection kits [20] [21]. All those advantageous characteristics make LAMP an ideal choice of on-site GMO detection method.…”

Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

“…nanotechnology-based sensors (Zhang et al 2005) or microfluidic systems, such as droplet -based , Chiou et al 2013, centrifugal (Cho et al 2007), paper (Martinez et al 2009, Fu et al 2010, Zuo et al 2013, capillary (T akayama et al 1999, Yager et al 2008, Gervais and Delamarche 2009, pneumatic (Legendre et al 2006, Mohan et al 2013 devices and lateral flow assays (Yager et al 2008, Chen et al 2010, You et al 2011). For the past decade, this has led to numerous publications about promising, early-stage technologies (Baeumner et al 2003, Leonard et al 2003, Kost 2006, Lazcka et al 2007, Lucchi et al 2010, Asiello and Baeumner 2011, Duarte et al 2011, Karlsson et al 2011, Schudel et al 2011, Matlock-Colangelo and Baeumner 2012, Lounsbury et al 2013, Mohan et al 2013. Despite some commercially available systems (e.g.…”

Development of a Fully Integrated Genetic Analysis Microsystem

“…[4] mechanical system)나 NEMS(Nano electro mechanical sys- Micro isothermal amplification system (NASBA). [7] Fig. 3.…”