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Room-temperature stable loop-mediated isothermal amplification (LAMP) reagents to detect leptospiral DNA

Room-temperature stable loop-mediated isothermal amplification (LAMP) reagents to detect... AbstractBackgroundLoop-mediated isothermal amplification (LAMP) is one of the most promising tools for rapidly detecting Leptospira spp. However, LAMP is hampered by cold storage to maintain the enzymatic activity of Bst DNA polymerase.ObjectiveTo overcome the drawback of cold storage requirement for LAMP reagents we modified the reagents by adding sucrose as stabilizer. We then sought to determine the stability at room temperature of the premixed LAMP reagents containing sucrose.MethodPremixed LAMP reagents with sucrose and without sucrose were prepared. The prepared mixtures were stored at room temperature for up to 60 days, and were subjected to LAMP reactions at various intervals using rat kidney samples to detect leptospiral DNA.ResultsThe premixed LAMP reagents with sucrose remained stable for 45 days while sucrose-free premixed LAMP reagents showed no amplification from day 1 of storage at room temperature up to day 14.ConclusionThe LAMP reagent system can be refined by using sucrose as stabilizer, thus allowing their storage at room temperature without the need for cold storage. The modified method enables greater feasibility of LAMP for field surveillance and epidemiology in resource-limited settings. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Asian Biomedicine de Gruyter

Room-temperature stable loop-mediated isothermal amplification (LAMP) reagents to detect leptospiral DNA

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References (19)

Publisher
de Gruyter
Copyright
© 2021 Pui-Yuei Lee et al., published by Sciendo
ISSN
1905-7415
eISSN
1875-855X
DOI
10.2478/abm-2021-0023
Publisher site
See Article on Publisher Site

Abstract

AbstractBackgroundLoop-mediated isothermal amplification (LAMP) is one of the most promising tools for rapidly detecting Leptospira spp. However, LAMP is hampered by cold storage to maintain the enzymatic activity of Bst DNA polymerase.ObjectiveTo overcome the drawback of cold storage requirement for LAMP reagents we modified the reagents by adding sucrose as stabilizer. We then sought to determine the stability at room temperature of the premixed LAMP reagents containing sucrose.MethodPremixed LAMP reagents with sucrose and without sucrose were prepared. The prepared mixtures were stored at room temperature for up to 60 days, and were subjected to LAMP reactions at various intervals using rat kidney samples to detect leptospiral DNA.ResultsThe premixed LAMP reagents with sucrose remained stable for 45 days while sucrose-free premixed LAMP reagents showed no amplification from day 1 of storage at room temperature up to day 14.ConclusionThe LAMP reagent system can be refined by using sucrose as stabilizer, thus allowing their storage at room temperature without the need for cold storage. The modified method enables greater feasibility of LAMP for field surveillance and epidemiology in resource-limited settings.

Journal

Asian Biomedicine – de Gruyter

Published: Aug 1, 2021

Keywords: leptospirosis; LAMP assay; temperature; room; stabilizer; sucrose

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