TY - JOUR
T1 - Rapid and sensitive multiplex single-tube nested PCR for the identification of five human Plasmodium species
AU - Saito, Takahiro
AU - Kikuchi, Aoi
AU - Kaneko, Akira
AU - Isozumi, Rie
AU - Teramoto, Isao
AU - Kimura, Masatsugu
AU - Hirasawa, Noriyasu
AU - Hiratsuka, Masahiro
N1 - Funding Information:
This work was supported by Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Number JP 16K15263 and AMED Translational Research Network Program. We would like to thank Evelyn Marie Gutierrez, Tohoku University, for carefully proofreading the manuscript.
Publisher Copyright:
© 2018 Elsevier B.V.
PY - 2018/6
Y1 - 2018/6
N2 - Malaria is caused by five species of Plasmodium in humans. Microscopy is currently used for pathogen detection, requiring considerable training and technical expertise as the parasites are often difficult to differentiate morphologically. Rapid diagnostic tests are as reliable as microscopy and offer faster diagnoses but possess lower detection limits and are incapable of distinguishing among the parasitic species. To improve global health efforts towards malaria control, a rapid, sensitive, species-specific, and economically viable diagnostic method is needed. In this study, we designed a malaria diagnostic method involving a multiplex single-tube nested PCR targeting Plasmodium mitochondrial cytochrome c oxidase III and single-stranded tag hybridization chromatographic printed-array strip. The detection sensitivity was found to be at least 40 times higher than that of agarose gel electrophoresis with ethidium bromide. This system also enables the identification of both single- and mixed-species malaria infections. The assay was validated with 152 Kenyan samples; using nested PCR as the standard, the assay's sensitivity and specificity were 88.7% and 100.0%, respectively. The turnaround time required, from PCR preparation to signal detection, is 90 min. Our method should improve the diagnostic speed, treatment efficacy, and control of malaria, in addition to facilitating surveillance within global malaria eradication programs.
AB - Malaria is caused by five species of Plasmodium in humans. Microscopy is currently used for pathogen detection, requiring considerable training and technical expertise as the parasites are often difficult to differentiate morphologically. Rapid diagnostic tests are as reliable as microscopy and offer faster diagnoses but possess lower detection limits and are incapable of distinguishing among the parasitic species. To improve global health efforts towards malaria control, a rapid, sensitive, species-specific, and economically viable diagnostic method is needed. In this study, we designed a malaria diagnostic method involving a multiplex single-tube nested PCR targeting Plasmodium mitochondrial cytochrome c oxidase III and single-stranded tag hybridization chromatographic printed-array strip. The detection sensitivity was found to be at least 40 times higher than that of agarose gel electrophoresis with ethidium bromide. This system also enables the identification of both single- and mixed-species malaria infections. The assay was validated with 152 Kenyan samples; using nested PCR as the standard, the assay's sensitivity and specificity were 88.7% and 100.0%, respectively. The turnaround time required, from PCR preparation to signal detection, is 90 min. Our method should improve the diagnostic speed, treatment efficacy, and control of malaria, in addition to facilitating surveillance within global malaria eradication programs.
KW - Diagnostic
KW - Malaria
KW - Multiplex nested PCR
KW - Plasmodium
KW - Printed-array strip
KW - Single-stranded tag hybridization
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U2 - 10.1016/j.parint.2018.01.005
DO - 10.1016/j.parint.2018.01.005
M3 - Article
C2 - 29374580
AN - SCOPUS:85041670844
VL - 67
SP - 277
EP - 283
JO - Parasitology International
JF - Parasitology International
SN - 1383-5769
IS - 3
ER -