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

Prudhvi Chand Mallepaddi Genomix Molecular Diagnostics Pvt. Ltd., Hyderabad, India;
Department of Biotechnology, Acharya Nagarjuna University, Guntur, India;

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Meng-Yee Lai Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia;

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Sudhakar Podha Department of Biotechnology, Acharya Nagarjuna University, Guntur, India;

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Choo-Huck Ooi Sarawak State Health Department, Jalan Diplomatik, Kuching, Malaysia

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Jonathan Wee-Kent Liew Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia;

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Rathnagiri Polavarapu Genomix Molecular Diagnostics Pvt. Ltd., Hyderabad, India;
Department of Biotechnology, Acharya Nagarjuna University, Guntur, India;

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Yee-Ling Lau Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia;

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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.

Author Notes

Address correspondence to Yee-Ling Lau, Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia. E-mail: lauyeeling@um.edu.my

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: prudhvi.mallepaddi@genomixbiotech.com and giri@genomixbiotech.com. 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: mengylai11@yahoo.com, jon_wkent@hotmail.com, and lauyeeling@um.edu.my. Sudhakar Podha, Department of Biotechnology, Acharya Nagarjuna University, Guntur, India, E-mail: sudhakarpodha@gmail.com. Choo-Huck Ooi, Sarawak State Health Department, Jalan Diplomatik, Kuching, Malaysia, E-mail: ooi.choo.huck@gmail.com.

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