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...
Main Authors: | , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2011-12-01
|
Series: | International Journal of Molecular Sciences |
Subjects: | |
Online Access: | http://www.mdpi.com/1422-0067/12/12/9108/ |
id |
doaj-3bc2639e3cc44045a4e400852cf71f17 |
---|---|
record_format |
Article |
spelling |
doaj-3bc2639e3cc44045a4e400852cf71f172020-11-25T00:20:27ZengMDPI AGInternational Journal of Molecular Sciences1422-00672011-12-0112129108912410.3390/ijms12129108Loop-Mediated Amplification Accelerated by Stem PrimersLaurence TisiGuy KiddleOlga GandelmanRebecca JacksonIsothermal 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.http://www.mdpi.com/1422-0067/12/12/9108/stem primersisothermal amplificationloop-mediated amplificationin vitro diagnosticsClostridium difficileListeria monocytogenesHIV |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Laurence Tisi Guy Kiddle Olga Gandelman Rebecca Jackson |
spellingShingle |
Laurence Tisi Guy Kiddle Olga Gandelman Rebecca Jackson Loop-Mediated Amplification Accelerated by Stem Primers International Journal of Molecular Sciences stem primers isothermal amplification loop-mediated amplification in vitro diagnostics Clostridium difficile Listeria monocytogenes HIV |
author_facet |
Laurence Tisi Guy Kiddle Olga Gandelman Rebecca Jackson |
author_sort |
Laurence Tisi |
title |
Loop-Mediated Amplification Accelerated by Stem Primers |
title_short |
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_sort |
loop-mediated amplification accelerated by stem primers |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2011-12-01 |
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. |
topic |
stem primers isothermal amplification loop-mediated amplification in vitro diagnostics Clostridium difficile Listeria monocytogenes HIV |
url |
http://www.mdpi.com/1422-0067/12/12/9108/ |
work_keys_str_mv |
AT laurencetisi loopmediatedamplificationacceleratedbystemprimers AT guykiddle loopmediatedamplificationacceleratedbystemprimers AT olgagandelman loopmediatedamplificationacceleratedbystemprimers AT rebeccajackson loopmediatedamplificationacceleratedbystemprimers |
_version_ |
1725367530550198272 |