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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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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. |
format | Online Article Text |
id | pubmed-6196555 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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