A comparison between Mini-loop mediated isothermal amplification and polymerase spiral reaction for selective amplification of short template DNA
Author(s)
Type
preprint
Abstract
<jats:title>Abstract</jats:title><jats:p>Isothermal amplification of circulating tumour-derived DNA (ctDNA) in the blood plasma could provide a rapid and cost effective alternative to PCR and NGS approaches for real-time cancer monitoring. Several variations of isothermal technologies exist, typically designed over unconstrained template length. Here, we compared the amplification efficiency of a compact loop mediated isothermal amplification reaction (termed AS-Mini-LAMP) with polymerase spiral reaction (PSR) suitable for analysis of ctDNA. Utilising 4-primers and targeting a 155 bp template encompassing the estrogen receptor (<jats:italic>ESR1)</jats:italic> c.1138G>C (p.E380Q) missense mutation.</jats:p><jats:p>Using synthetic E380Q template DNA and Bst2.0 polymerase, results demonstrate that AS-Mini-LAMP was capable of selective mutant allele DNA amplification to a limit of 1,000 mutant copies, whereas no specific amplification was observed by PSR. The alternative use of Bst3.0 polymerase for either AS-Mini-LAMP or PSR revealed non-canonical events that underpin potentially misleading results when employing isothermal chemistries. In conclusion, AS-Mini-LAMP is more suited to mutation detection than PSR.</jats:p>
Date Issued
2022-05-19
Date Acceptance
2022-05-20
Citation
bioRxiv
Publisher
Nature Publishing Group
Journal / Book Title
bioRxiv
Copyright Statement
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
License URL
Publication Status
Published