Abstract

Nucleic acid amplification technologies (NAATs) offer the most sensitive tests in the clinical laboratory. These techniques are used as a powerful tool for screening and diagnosis of infectious diseases. Isothermal methods, as an alternative to polymerase chain reaction (PCR), require no thermocycling machine and can mostly be performed with reduced time, high throughput, and accurate and reliable results. However, current molecular diagnostic approaches generally need manual analysis by qualified and experienced personal which is a highly complex, time-consuming and labor-intensive task. Thus, the demand for simpler, miniaturized systems and assays for pathogen detection is steadily increasing. Microfluidic platforms and lab-on-a-chip devices have many advantages such as small sample volume, portability and rapid detection time and enable point-of-care diagnosis. In this article, we review several isothermal amplification methods and their implementation in microsystems in relation to quantification of nucleic acids.

Highlights

  • The idea of miniaturization of available technologies was created by Richard P

  • We review several isothermal amplification methods and their implementation in microsystems in relation to quantification of nucleic acids

  • In 1982, Petersen described the entire potential of microsystem technology, and in the following years and decades a wealth of concepts for microsystems, possible applications and new miniaturized systems were developed by today [2]

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Summary

Introduction

The idea of miniaturization of available technologies was created by Richard P. Miniaturized systems for the analysis of genetic sequences are a major cornerstone for the development and marketability of pointof-care (POC) devices [6]. These devices can be characterized by a high integration of several functionalities onto a single platform, e.g. cumbersome lab processes like sample preparation, nucleic acid amplification and detection. Microsystems lead to portable, easy-to-use devices, which might be applied by lay users These systems have shown that the analysis of samples can be moved from a specialized laboratory closer to the patient, aiming at POC, or even to in-field testing [8,9]. Comprehensive references will direct the reader to the more detailed descriptions of the biology behind the amplification reactions

Strategies to Miniaturize Isothermal Amplification Reactions
Lateral flow dipstick
SYBR green I fluorescence
The first integration of a NASBA reaction into a microfluidic
SiHa cell line
Cellular and molecular analysis platform realized by an integrated
Visual detection
Fluorescence image
Few cells
Various other methods
Findings
Conclusion
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