Abstract
The easy and rapid spread of bacterial contamination and the risk it poses to human health makes evident the need for analytical methods alternative to conventional time-consuming laboratory-based techniques for bacterial detection. To tackle this demand, biosensors based on isothermal DNA amplification methods have emerged, which avoid the need for thermal cycling, thus facilitating their integration into small and low-cost devices for in situ monitoring. This review focuses on the breakthroughs made on biosensors based on isothermal amplification methods for the detection of bacteria in the field of food safety and environmental monitoring. Optical and electrochemical biosensors based on loop mediated isothermal amplification (LAMP), rolling circle amplification (RCA), recombinase polymerase amplification (RPA), helicase dependent amplification (HDA), strand displacement amplification (SDA), and isothermal strand displacement polymerisation (ISDPR) are described, and an overview of their current advantages and limitations is provided. Although further efforts are required to harness the potential of these emerging analytical techniques, the coalescence of the different isothermal amplification techniques with the wide variety of biosensing detection strategies provides multiple possibilities for the efficient detection of bacteria far beyond the laboratory bench.
Highlights
Bacterial contamination of food, water and environment is a crucial safety issue as it is linked to human and animal diseases, some of them leading to death and great economic losses
Since the molecular switch used in the strand displacement amplification (SDA)-based biosensor was not related to the aptamer sequence, the same strategy was applied to the detection of E. coli by changing the aptamer, the terminals sequence of the complementary single-stranded DNA (ssDNA) and the primers
Current reduction increased as the target strand was replaced blue away from electrode surface and resulted in a decrease bymolecules the amplified strand andthe it was hybridised to another hairpin probe, achieving of an the LODcurrent response
Summary
Water and environment is a crucial safety issue as it is linked to human and animal diseases, some of them leading to death and great economic losses. Unlike typical thermal cycling amplification, isothermal techniques are carried out at a constant temperature, which reduces the power needed and makes them more compatible for integration into miniaturised systems [4]. These techniques have provided biosensors with the relevant sensitivity and specificity for the detection of different analytes such as microalgae [6,7]. The objective of this work is to provide a comprehensive overview of the biosensors based on isothermal DNA amplification methods for bacterial detection with application in the fields of food safety and environmental monitoring, affecting human, animal and plant health. It faces the key factors that are crucial in the development of these biosensors and the advantages and limitations of the use of isothermal DNA amplification methods
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