Sensitive detection of Plasmodium vivax using a high-throughput, colourimetric loop mediated isothermal amplification (HtLAMP) platform: a potential novel tool for malaria elimination

Britton, S. et al. (2016) Sensitive detection of Plasmodium vivax using a high-throughput, colourimetric loop mediated isothermal amplification (HtLAMP) platform: a potential novel tool for malaria elimination. PLoS Neglected Tropical Diseases, 10(2), e0004443. (doi: 10.1371/journal.pntd.0004443) (PMID:26870958) (PMCID:PMC4752294)

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Abstract

Introduction: Plasmodium vivax malaria has a wide geographic distribution and poses challenges to malaria elimination that are likely to be greater than those of P. falciparum. Diagnostic tools for P. vivax infection in non-reference laboratory settings are limited to microscopy and rapid diagnostic tests but these are unreliable at low parasitemia. The development and validation of a high-throughput and sensitive assay for P. vivax is a priority. Methods: A high-throughput LAMP assay targeting a P. vivax mitochondrial gene and deploying colorimetric detection in a 96-well plate format was developed and evaluated in the laboratory. Diagnostic accuracy was compared against microscopy, antigen detection tests and PCR and validated in samples from malaria patients and community controls in a district hospital setting in Sabah, Malaysia. Results: The high throughput LAMP-P. vivax assay (HtLAMP-Pv) performed with an estimated limit of detection of 1.4 parasites/ μL. Assay primers demonstrated cross-reactivity with P. knowlesi but not with other Plasmodium spp. Field testing of HtLAMP-Pv was conducted using 149 samples from symptomatic malaria patients (64 P. vivax, 17 P. falciparum, 56 P. knowlesi, 7 P. malariae, 1 mixed P. knowlesi/P. vivax, with 4 excluded). When compared against multiplex PCR, HtLAMP-Pv demonstrated a sensitivity for P. vivax of 95% (95% CI 87–99%); 61/64), and specificity of 100% (95% CI 86–100%); 25/25) when P. knowlesi samples were excluded. HtLAMP-Pv testing of 112 samples from asymptomatic community controls, 7 of which had submicroscopic P. vivax infections by PCR, showed a sensitivity of 71% (95% CI 29–96%; 5/7) and specificity of 93% (95% CI87-97%; 98/105). Conclusion: This novel HtLAMP-P. vivax assay has the potential to be a useful field applicable molecular diagnostic test for P. vivax infection in elimination settings.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Fornace, Dr Kimberly
Authors: Britton, S., Cheng, Q., Grigg, M. J., Poole, C. B., Pasay, C., William, T., Fornace, K., Anstey, N. M., Sutherland, C. J., Drakeley, C., and McCarthy, J. S.
College/School:College of Medical Veterinary and Life Sciences > Institute of Biodiversity Animal Health and Comparative Medicine
College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine
Journal Name:PLoS Neglected Tropical Diseases
Publisher:Public Library of Science
ISSN:1935-2727
ISSN (Online):1935-2735
Published Online:12 February 2016
Copyright Holders:Copyright © 2016 Britton et al.
First Published:First published in PLoS Neglected Tropical Diseases 10(2): e0004443
Publisher Policy:Reproduced under a Creative Commons License

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