<abstract xmlns="http://www.w3.org/1999/xhtml">

<sec><h3>Background</h3>
<p>Loop-mediated isothermal amplification (LAMP) is one of the most promising tools for rapidly detecting <italic>Leptospira</italic> spp. However, LAMP is hampered by cold storage to maintain the enzymatic activity of <italic>Bst</italic> DNA polymerase.</p>
</sec>
<sec><h3>Objective</h3>
<p>To 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.</p>
</sec>
<sec><h3>Method</h3>
<p>Premixed 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.</p>
</sec>
<sec><h3>Results</h3>
<p>The 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.</p>
</sec>
<sec><h3>Conclusion</h3>
<p>The 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.</p>
</sec>
</abstract>

Background
Loop-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.

Objective
To 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.

Method
Premixed 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.

Results
The 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.

Conclusion
The 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.

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

Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health Sciences

Asian Biomedicine

This work is licensed under the Creative Commons Attribution 4.0 International License.

Background
Loop-mediated isothermal amplification (LAMP) is one of the most promising tools for rapidly detecting Leptospira spp. However, LAMP is hampered by...

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

Article Category: Technical report

Pubblicato online: 20 ago 2021

Pagine: 183 - 189

DOI: https://doi.org/10.2478/abm-2021-0023

Parole chiave
leptospirosis, LAMP assay, temperature, room, stabilizer, sucrose

© 2021 Pui-Yuei Lee et al., published by Sciendo

This work is licensed under the Creative Commons Attribution 4.0 International License.

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

Article Category: Technical report

Pubblicato online: 20 ago 2021

Pagine: 183 - 189

DOI: https://doi.org/10.2478/abm-2021-0023

Parole chiaveleptospirosis, LAMP assay, temperature, room, stabilizer, sucrose

© 2021 Pui-Yuei Lee et al., published by Sciendo

This work is licensed under the Creative Commons Attribution 4.0 International License.

Parole chiave
leptospirosis, LAMP assay, temperature, room, stabilizer, sucrose