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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 amidite-labeled loop primers to the LAMP reaction solution, the end product can be visualized on a LFD. The entire procedure takes approximately 42 minutes to complete, LAMP assay exhibited high sensitivity, as the detection limit was 0.01 pg/μL for all five Plasmodium species. It was 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 represents a robust approach with potential suitability for use in resource-constrained laboratories. We believe that LAMP-LFD has a potential to be developed as point-of-care diagnostic tool in future.
These authors contributed equally to this work.
Financial support: This study was supported by University Malaya PPP Research Grant (PG066-2016A) and Department of Biotechnology under SIBRI Project on Malarial Diagnostics (File no: BT/SBIRI/358/57-B6/2007).
Authors’ addresses: Prudhvi Chand Mallepaddi and Rathnagiri Polavarapu, Genomix Molecular Diagnostics Pvt. Ltd., Hyderabad, India, E-mails: email@example.com and firstname.lastname@example.org. Meng-Yee Lai, Jonathan Wee-Kent Liew, and Yee-Ling Lau, Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia, E-mails: email@example.com, firstname.lastname@example.org, and email@example.com. Sudhakar Podha, Department of Biotechnology, Acharya Nagarjuna University, Guntur, India, E-mail: firstname.lastname@example.org. Choo-Huck Ooi, Sarawak State Health Department, Jalan Diplomatik, Kuching, Malaysia, E-mail: email@example.com.