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Loop-Mediated Amplification Accelerated by Stem Primers

Isothermal nucleic acid amplifications (iNAATs) have become an important alternative to PCR for in vitro molecular diagnostics in all fields. Amongst iNAATs Loop-mediated amplification (LAMP) has gained much attention over the last decade because of the simplicity of hardware requirements. LAMP demo...

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Detalles Bibliográficos
Autores principales: Gandelman, Olga, Jackson, Rebecca, Kiddle, Guy, Tisi, Laurence
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3257119/
https://www.ncbi.nlm.nih.gov/pubmed/22272122
http://dx.doi.org/10.3390/ijms12129108
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author Gandelman, Olga
Jackson, Rebecca
Kiddle, Guy
Tisi, Laurence
author_facet Gandelman, Olga
Jackson, Rebecca
Kiddle, Guy
Tisi, Laurence
author_sort Gandelman, Olga
collection PubMed
description Isothermal nucleic acid amplifications (iNAATs) have become an important alternative to PCR for in vitro molecular diagnostics in all fields. Amongst iNAATs Loop-mediated amplification (LAMP) has gained much attention over the last decade because of the simplicity of hardware requirements. LAMP demonstrates performance equivalent to that of PCR, but its application has been limited by the challenging primer design. The design of six primers in LAMP requires a selection of eight priming sites with significant restrictions imposed on their respective positioning and orientation. In order to relieve primer design constraints we propose an alternative approach which uses Stem primers instead of Loop primers and demonstrate the application of STEM-LAMP in assaying for Clostridium difficile, Listeria monocytogenes and HIV. Stem primers used in LAMP in combination with loop-generating and displacement primers gave significant benefits in speed and sensitivity, similar to those offered by Loop primers, while offering additional options of forward and reverse orientations, multiplexing, use in conjunction with Loop primers or even omission of one or two displacement primers, where necessary. Stem primers represent a valuable alternative to Loop primers and an additional tool for IVD assay development by offering more choices for primer design at the same time increasing assay speed, sensitivity, and reproducibility.
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spelling pubmed-32571192012-01-23 Loop-Mediated Amplification Accelerated by Stem Primers Gandelman, Olga Jackson, Rebecca Kiddle, Guy Tisi, Laurence Int J Mol Sci Article Isothermal nucleic acid amplifications (iNAATs) have become an important alternative to PCR for in vitro molecular diagnostics in all fields. Amongst iNAATs Loop-mediated amplification (LAMP) has gained much attention over the last decade because of the simplicity of hardware requirements. LAMP demonstrates performance equivalent to that of PCR, but its application has been limited by the challenging primer design. The design of six primers in LAMP requires a selection of eight priming sites with significant restrictions imposed on their respective positioning and orientation. In order to relieve primer design constraints we propose an alternative approach which uses Stem primers instead of Loop primers and demonstrate the application of STEM-LAMP in assaying for Clostridium difficile, Listeria monocytogenes and HIV. Stem primers used in LAMP in combination with loop-generating and displacement primers gave significant benefits in speed and sensitivity, similar to those offered by Loop primers, while offering additional options of forward and reverse orientations, multiplexing, use in conjunction with Loop primers or even omission of one or two displacement primers, where necessary. Stem primers represent a valuable alternative to Loop primers and an additional tool for IVD assay development by offering more choices for primer design at the same time increasing assay speed, sensitivity, and reproducibility. Molecular Diversity Preservation International (MDPI) 2011-12-08 /pmc/articles/PMC3257119/ /pubmed/22272122 http://dx.doi.org/10.3390/ijms12129108 Text en © 2011 by the authors; licensee MDPI, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0 This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Gandelman, Olga
Jackson, Rebecca
Kiddle, Guy
Tisi, Laurence
Loop-Mediated Amplification Accelerated by Stem Primers
title Loop-Mediated Amplification Accelerated by Stem Primers
title_full Loop-Mediated Amplification Accelerated by Stem Primers
title_fullStr Loop-Mediated Amplification Accelerated by Stem Primers
title_full_unstemmed Loop-Mediated Amplification Accelerated by Stem Primers
title_short Loop-Mediated Amplification Accelerated by Stem Primers
title_sort loop-mediated amplification accelerated by stem primers
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3257119/
https://www.ncbi.nlm.nih.gov/pubmed/22272122
http://dx.doi.org/10.3390/ijms12129108
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