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

Britton, Sumudu, Cheng, Qin, Grigg, Matthew J., Poole, Catherine B., Pasay, Cielo, William, Timothy, Fornace, Kimberley, Anstey, Nicholas M., Sutherland, Colin J., Drakeley, Chris and McCarthy, James S. (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: . doi:10.1371/journal.pntd.0004443


Author Britton, Sumudu
Cheng, Qin
Grigg, Matthew J.
Poole, Catherine B.
Pasay, Cielo
William, Timothy
Fornace, Kimberley
Anstey, Nicholas M.
Sutherland, Colin J.
Drakeley, Chris
McCarthy, James S.
Title Sensitive detection of Plasmodium vivax using a high-throughput, colourimetric loop mediated isothermal amplification (HtLAMP) platform: a potential novel tool for malaria elimination
Formatted title
Sensitive detection of Plasmodium vivax using a high-throughput, colourimetric loop mediated isothermal amplification (HtLAMP) platform: a potential novel tool for malaria elimination
Journal name PLoS Neglected Tropical Diseases   Check publisher's open access policy
ISSN 1935-2735
1935-2727
Publication date 2016-02-12
Year available 2016
Sub-type Article (original research)
DOI 10.1371/journal.pntd.0004443
Open Access Status DOI
Volume 10
Issue 2
Total pages 16
Place of publication San Francisco, CA, United States
Publisher Public Library of Science
Collection year 2017
Language eng
Formatted 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.
Keyword Plasmodium vivax
P. vivax
High-Throughput Colourimetric Loop Mediated Isothermal Amplification (HtLAMP)
Malaria elimination
Human malaria
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
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