Sensitive Detection of Plasmodium Vivax Using a High-Throughput, Colourimetric Loop Mediated Isothermal Amplification (HtLAMP) Platform: A Potential Novel Tool for Malaria Elimination

Overview

Journal PLoS Negl Trop Dis

Specialties Infectious Diseases
Tropical Medicine

Date 2016 Feb 13

PMID 26870958

Citations 31

Authors

Sumudu Britton

Qin Cheng

Matthew J Grigg

Catherine B Poole

Cielo Pasay

Timothy William

Kimberley Fornace

Nicholas M Anstey

Colin J Sutherland

Chris Drakeley

James S McCarthy

Affiliations

Soon will be listed here.

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.

Citing Articles

: What Should We Know?.

Muh F, Erwina A, Fitriana F, Syahada J, Cahya A, Choe S Microorganisms. 2024; 12(8).

PMID: 39203449 PMC: 11356028. DOI: 10.3390/microorganisms12081607.

Recombinase-Aided Loop-Mediated Isothermal Amplification on Human Plasmodium knowlesi.

Lai M, Abdul Hamid M, Jelip J, Mudin R, Lau Y Am J Trop Med Hyg. 2024; 110(4):648-652.

PMID: 38412548 PMC: 10993835. DOI: 10.4269/ajtmh.23-0572.

Is qPCR always the most sensitive method for malaria diagnostic quality surveillance?.

Koepfli C Malar J. 2023; 22(1):380.

PMID: 38102649 PMC: 10722660. DOI: 10.1186/s12936-023-04822-w.

Malaria diagnosis using a combined system of a simple and fast extraction method with a lyophilised Dual-LAMP assay in a non-endemic setting.

Martin Ramirez A, Baron Argos L, Lanza Suarez M, Carmona Rubio C, Perez-Ayala A, Hisam S Pathog Glob Health. 2023; 118(1):80-90.

PMID: 37415348 PMC: 10769111. DOI: 10.1080/20477724.2023.2232595.

Sensitive Detection of Asymptomatic and Symptomatic Malaria with Seven Novel Parasite-Specific LAMP Assays and Translation for Use at Point-of-Care.

Malpartida-Cardenas K, Moser N, Ansah F, Pennisi I, Prah D, Amoah L Microbiol Spectr. 2023; 11(3):e0522222.

PMID: 37158750 PMC: 10269850. DOI: 10.1128/spectrum.05222-22.

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