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Combination of PURE-DNA extraction and LAMP-DNA amplification methods for accurate malaria diagnosis on dried blood spots

BACKGROUND: Malaria is one of the most important parasitic infectious diseases for which almost half of the world’s population is at risk. Although several diagnostic methods are now available to detect the infection, more sensitive and applicable tests are still required in the field. The loop-medi...

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Autores principales: Vincent, Jeanne Perpétue, Komaki-Yasuda, Kanako, Iwagami, Moritoshi, Kawai, Satoru, Kano, Shigeyuki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6196555/
https://www.ncbi.nlm.nih.gov/pubmed/30348162
http://dx.doi.org/10.1186/s12936-018-2527-7
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author Vincent, Jeanne Perpétue
Komaki-Yasuda, Kanako
Iwagami, Moritoshi
Kawai, Satoru
Kano, Shigeyuki
author_facet Vincent, Jeanne Perpétue
Komaki-Yasuda, Kanako
Iwagami, Moritoshi
Kawai, Satoru
Kano, Shigeyuki
author_sort Vincent, Jeanne Perpétue
collection PubMed
description BACKGROUND: Malaria is one of the most important parasitic infectious diseases for which almost half of the world’s population is at risk. Although several diagnostic methods are now available to detect the infection, more sensitive and applicable tests are still required in the field. The loop-mediated isothermal amplification (LAMP) method is a DNA amplification tool in which the DNA amplification can be achieved by incubation at a stable temperature. A malaria detection kit based on this methodology has already been commercialized and is being used in some countries. The kit includes two reaction tubes: one targeting the common Plasmodium genus (Pan tube) and the other specifically targeting Plasmodium falciparum (Pf tube). In parallel, a simple DNA extraction method, the procedure for ultra rapid extraction (PURE), which can produce a DNA solution suitable for the LAMP reaction without the use of a centrifuge, has also become available. In this study, the sensitivity of the combination of the PURE and LAMP methods (PURE–LAMP) was evaluated with archived dried clinical blood samples of imported malaria cases, including P. falciparum, Plasmodium vivax, Plasmodium ovale, and Plasmodium malariae. RESULTS: Using a nested PCR as the reference, 117 samples including 46 P. falciparum, 7 P. vivax, 9 P. ovale, 4 P. malariae, and 51 negative cases were tested. The PURE–LAMP Pan correctly identified 64 of the 66 positives and the 51 negatives. Among the Pan-positive samples 45 P. falciparum were also detected with the PURE–LAMP Pf. The PURE–LAMP Pan and PURE–LAMP Pf had respective sensitivities of 96.96% (95% CI 89.47–99.63) and 97.82% (95% CI 88.47–99.94) and common specificity of 1. CONCLUSION: The PURE–LAMP system is accurate when used with dried blood spots and extendable to the field. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12936-018-2527-7) contains supplementary material, which is available to authorized users.
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spelling pubmed-61965552018-10-30 Combination of PURE-DNA extraction and LAMP-DNA amplification methods for accurate malaria diagnosis on dried blood spots Vincent, Jeanne Perpétue Komaki-Yasuda, Kanako Iwagami, Moritoshi Kawai, Satoru Kano, Shigeyuki Malar J Research BACKGROUND: Malaria is one of the most important parasitic infectious diseases for which almost half of the world’s population is at risk. Although several diagnostic methods are now available to detect the infection, more sensitive and applicable tests are still required in the field. The loop-mediated isothermal amplification (LAMP) method is a DNA amplification tool in which the DNA amplification can be achieved by incubation at a stable temperature. A malaria detection kit based on this methodology has already been commercialized and is being used in some countries. The kit includes two reaction tubes: one targeting the common Plasmodium genus (Pan tube) and the other specifically targeting Plasmodium falciparum (Pf tube). In parallel, a simple DNA extraction method, the procedure for ultra rapid extraction (PURE), which can produce a DNA solution suitable for the LAMP reaction without the use of a centrifuge, has also become available. In this study, the sensitivity of the combination of the PURE and LAMP methods (PURE–LAMP) was evaluated with archived dried clinical blood samples of imported malaria cases, including P. falciparum, Plasmodium vivax, Plasmodium ovale, and Plasmodium malariae. RESULTS: Using a nested PCR as the reference, 117 samples including 46 P. falciparum, 7 P. vivax, 9 P. ovale, 4 P. malariae, and 51 negative cases were tested. The PURE–LAMP Pan correctly identified 64 of the 66 positives and the 51 negatives. Among the Pan-positive samples 45 P. falciparum were also detected with the PURE–LAMP Pf. The PURE–LAMP Pan and PURE–LAMP Pf had respective sensitivities of 96.96% (95% CI 89.47–99.63) and 97.82% (95% CI 88.47–99.94) and common specificity of 1. CONCLUSION: The PURE–LAMP system is accurate when used with dried blood spots and extendable to the field. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12936-018-2527-7) contains supplementary material, which is available to authorized users. BioMed Central 2018-10-22 /pmc/articles/PMC6196555/ /pubmed/30348162 http://dx.doi.org/10.1186/s12936-018-2527-7 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Vincent, Jeanne Perpétue
Komaki-Yasuda, Kanako
Iwagami, Moritoshi
Kawai, Satoru
Kano, Shigeyuki
Combination of PURE-DNA extraction and LAMP-DNA amplification methods for accurate malaria diagnosis on dried blood spots
title Combination of PURE-DNA extraction and LAMP-DNA amplification methods for accurate malaria diagnosis on dried blood spots
title_full Combination of PURE-DNA extraction and LAMP-DNA amplification methods for accurate malaria diagnosis on dried blood spots
title_fullStr Combination of PURE-DNA extraction and LAMP-DNA amplification methods for accurate malaria diagnosis on dried blood spots
title_full_unstemmed Combination of PURE-DNA extraction and LAMP-DNA amplification methods for accurate malaria diagnosis on dried blood spots
title_short Combination of PURE-DNA extraction and LAMP-DNA amplification methods for accurate malaria diagnosis on dried blood spots
title_sort combination of pure-dna extraction and lamp-dna amplification methods for accurate malaria diagnosis on dried blood spots
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6196555/
https://www.ncbi.nlm.nih.gov/pubmed/30348162
http://dx.doi.org/10.1186/s12936-018-2527-7
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