Naked‐eye detection strategies coupled with isothermal nucleic acid amplification techniques for the detection of human pathogens

Abstract

Nucleic acid amplification‐based techniques have gained acceptance by the scientific, and general, community as reference methodologies for many different applications. Since the development of the gold standard of these techniques, polymerase chain reaction (PCR), back in the 1980s many improvements have been made, and alternative techniques emerged reporting improvements over PCR. Among these, isothermal amplification approaches resulted of particular interest as could overcome the need of specialized equipment to accurately control temperature changes, but it was after year 2000 that these techniques have flourished in a huge number of novel alternatives with many different degrees of complexities and requirements. An added value is their possibility to be combined with many different naked‐eye detection strategies, simplifying the resources needed, allowing to reduce cost, and serving as the basis for novel developments of lab‐on‐chip systems, and miniaturized devices, for point‐of‐care testing. In this review, we will go over different types of naked‐eye detection strategies, combined with isothermal amplification. This will provide the readers up‐to‐date information for them to select the most appropriate strategies depending on the particular needs and resources for their experimental setup.

Application of MinION sequencing as a tool for the rapid detection and characterization of Listeria monocytogenes in smoked salmon

“…Even though many different molecular methods, and detection platforms, have been developed in recent years (El Sheikha et al, 2018 ; El Sheikha, 2021 ; Garrido-Maestu and Prado, 2022 ), when it comes to the detection of foodborne pathogens, which typically are present in foods at very low concentrations, some kind of sample treatment is always needed (Brehm Stecher et al, 2009 ). The classical way to proceed is to culture the microorganisms in a suitable broth.…”

Microfluidic-based colorimetric nucleic acid detection of pathogens

“…32 Some significant advantages of colorimetric readouts include ease of interpretation, label-free signal transduction, cost-effectiveness, and simplicity. 33 The color sensing schemes when coupled with nucleic acid detection assays for pathogens, are broadly categorized into (a) pH responsive colorimetric change, 34 (b) color-change via chromogenic indicators, 35 (c) colorimetric or florescence change via intercalating dyes, 36 (d) nanoparticle mediated colorimetric change. 37 One notable obstacle with colorimetric readout is that the human eye is insensitive to low level color contrasts; thus, discriminating outputs between distinct samples is difficult with the naked eye since most conventional colorimetric approaches primarily depend on the colored output's optical density variation.…”

A DNA walker triggered isothermal amplification method based on freezing construction of AuNP probes and its application in ricin detection

“…19,20 Due to its remarkable signal amplification effect, isothermal amplification of nucleic acids has emerged as a promising alternative to polymerase chain reaction (PCR). 21,22 Isothermal amplification is a simple process that rapidly and efficiently accumulates nucleic acid at a constant temperature for applications in bioanalysis, diagnostics, nanotechnology, and materials science. 23,24 Several isothermal amplification techniques such as rolling circle amplification (RCA), exponential amplification reaction (EXPAR), strand displacement amplification (SDA), hybridization chain reaction (HCR), and DNAzyme have been proposed.…”