Development of Loop-Mediated Isothermal Amplification–Based Lateral Flow Device Method for the Detection of Malaria

Document Type

Article

Publication Date

1-1-2018

Abstract

The present study aims to develop a method for rapid diagnosis of malaria using loop-mediated isothermal amplification (LAMP) combined with a lateral flow device (LFD). By adding the biotin-labeled and fluorescein amiditelabeled loop primers to the LAMP reaction solution, the end product can be visualized on a LFD. The entire procedure takes approximately 42 minutes tocomplete, LAMP assay exhibited high sensitivity, asthe detection limit was 0.01 pg/μL for all five Plasmodium species. Itwas demonstrated that all Plasmodium knowlesi (N = 90) and Plasmodium vivax(N= 56) were positively amplified by LAMP-LFD assay, whereas healthy donor samples (N = 8) were negative. However, not all mixed infections were positive, and other infected nonmalaria samples were negative. Loop-mediated isothermal amplification-LFD representsarobust approach with potential suitability for useinresource-constrained laboratories. We believe that LAMP-LFD has a potential to be developed as point-of-care diagnostic tool in future.

Keywords

controlled study, lateral flow device, limit of detection, loop mediated isothermal amplification, malaria, mixed infection, nonhuman, Plasmodium knowlesi, Plasmodium vivax, point of care testing, sensitivity analysis

Divisions

fac_med

Funders

University Malaya PPP Research Grant (PG066-2016A),Department of Biotechnology under SIBRI Project on Malarial Diagnostics (File no: BT/ SBIRI/358/57-B6/2007)

Publication Title

The American Journal of Tropical Medicine and Hygiene

Volume

99

Issue

3

Publisher

American Society of Tropical Medicine and Hygiene

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