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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.

plos neglected tropical diseases

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