TY - JOUR
T1 - Sensitive Detection of Plasmodium vivax Using a High-Throughput, Colourimetric Loop Mediated Isothermal Amplification (HtLAMP) Platform
T2 - A Potential Novel Tool for Malaria Elimination
AU - Britton, S
AU - Cheng, Q
AU - Grigg, Matthew
AU - Poole, CB
AU - Pasay, Cielo
AU - William, Timothy
AU - Fornace, K
AU - Anstey, Nicholas
AU - Sutherland, Colin
AU - Drakeley, Chris
AU - McCarthy, James
N1 - Australian National Health and Medical Research Council (Program grant #1037304, Project Grant #1045156
PY - 2016
Y1 - 2016
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84959290432&partnerID=8YFLogxK
U2 - 10.1371/journal.pntd.0004443
DO - 10.1371/journal.pntd.0004443
M3 - Article
C2 - 26870958
VL - 10
SP - 1
EP - 16
JO - PLoS Neglected Tropical Diseases
JF - PLoS Neglected Tropical Diseases
SN - 1935-2727
IS - 2
ER -