Issue 6, 2024

Effects of single and multiple nucleotide mutations on loop-mediated isothermal amplification

Abstract

Testing is pivotal for early identification of disease and subsequent infection control. Pathogens’ nucleic acid sequence can change due to naturally-occurring genetic drift or intentional modification. Because of the reliance on molecular assays for human, animal, and plant disease diagnosis, we must understand how nucleotide mutations affect test accuracy. Primers designed against original lineages of a pathogen may be less efficient at detecting variants with genetic changes in priming regions. Here, we made single- and multi-point mutations in priming regions of a model SARS-CoV-2 template that was used as input for a loop-mediated isothermal amplification (LAMP) assay. We found that many of the modifications impacted assay sensitivity, amplification speed, or both. Further research exploring mutations at every position in each of the eight priming regions should be conducted to evaluate trends and determine generalizability.

Graphical abstract: Effects of single and multiple nucleotide mutations on loop-mediated isothermal amplification

Supplementary files

Article information

Article type
Communication
Submitted
06 Nov 2023
Accepted
13 Feb 2024
First published
21 Feb 2024
This article is Open Access
Creative Commons BY-NC license

Analyst, 2024,149, 1701-1708

Effects of single and multiple nucleotide mutations on loop-mediated isothermal amplification

T. J. Moehling, E. R. Browne and R. J. Meagher, Analyst, 2024, 149, 1701 DOI: 10.1039/D3AN01927F

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