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July 29, 2024
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Post in: Application Notes, Molecular Diagnostics
Polaris® BstY: Optimizing Speed and Specificity in LAMP assays
Rita S. Simões¹´² João S. Teodoro¹ & Carlos M.G.A. Fontes¹
¹ Department of Research and Development, NZYtech, Lisbon, Portugal; [2] CIISA, Faculty of Veterinary Medicine, University of Lisbon, Portugal
* rita.simoes@nzytech.com
Introduction
Strand-displacing polymerases are a critical element in loop-mediated isothermal amplification (LAMP), in which the lack of thermal cycling reduces equipment needs and improves the time to result, especially for point-of-care (POC) applications[1, 2].
Although LAMP assays offer high target specificity, the complex reaction matrix which involves up to six primer sequences and a relatively low reaction temperature (55-65 °C) often leads to non-specific amplification[3]. While Bst DNA polymerase remains the enzyme of choice for LAMP reactions[4], its relatively low reaction temperature limits its ability to perform DNA amplification at higher temperatures, unlike other polymerases used in various molecular diagnostics techniques, such as Taq DNA polymerase[5].
To address these issues, NZYtech developed Polaris® BstY Polymerase: a variant of wild-type enzyme with remarkable thermal resistance. Engineered for high-temperature LAMP assays (optimal activity at 68-69 °C), Polaris® BstY Polymerase surpasses traditional enzymes by operating effectively between 65 and 70 °C. This not only increases the specificity of the assay, by reducing non-specific amplifications, but also mitigates the effects of inhibitors present in various sample types, essential traits for both clinical and research applications. Resistance to inhibitors is particularly relevant in POC applications, where samples are typically unprocessed or subjected to very simple extraction procedures.
In this application note, we detail and characterize the potential of Polaris® BstY Polymerase as an indispensable ally in the pursuit of precise, reliable LAMP diagnostics across various conditions. Polaris® BstY Polymerase is available in three formats: high concentration without glycerol, the standard glycerol format, and a lyophilized option, as detailed on the last page of this publication.
