Phosphorothioated Primers Lead to Loop-Mediated Isothermal Amplification at Low Temperatures

Loop-mediated isothermal amplification (LAMP) is an extremely powerful tool for the detection of nucleic acids with high sensitivity and specificity. However, LAMP shows optimal performance at around 65 °C, which limits applications in point-of-care-testing (POCT). Here, we have developed a version...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2018-07, Vol.90 (14), p.8290-8294
Main Authors: Cai, Sheng, Jung, Cheulhee, Bhadra, Sanchita, Ellington, Andrew D
Format: Article
Language:English
Subjects:
Analytical chemistry
Chemistry
Concatamers
Deoxyribonucleic acid
DNA
Low temperature
Low temperature physics
Nucleic acids
Phosphorus
Primers
Proteins
Sensitivity
Sensors
Single-stranded DNA
Urea
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Summary:Loop-mediated isothermal amplification (LAMP) is an extremely powerful tool for the detection of nucleic acids with high sensitivity and specificity. However, LAMP shows optimal performance at around 65 °C, which limits applications in point-of-care-testing (POCT). Here, we have developed a version of LAMP that uses phosphorothioated primers (PS-LAMP) to enable more efficient hairpin formation and extension at the termini of growing concatamers, and that therefore works at much lower temperatures. By including additional factors such as chaotropes (urea) and single-stranded DNA binding protein (SSB), the sensitivities and selectivities for amplicon detection with PS-LAMP at 40 °C were comparable with a regular LAMP reaction at 65 °C.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.8b02062