Development of a Multiplex Loop-Mediated Isothermal Amplification Assay for Diagnosis of Plasmodium spp., Plasmodium falciparum and Plasmodium vivax

Malaria, caused by the parasite Plasmodium and transmitted by mosquitoes, is an epidemic that mainly occurs in tropical and subtropical regions. As treatments differ across species of malarial parasites, there is a need to develop rapid diagnostic methods to differentiate malarial...

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Autores principales:	Woong Sik Jang, Da Hye Lim, YoungLan Choe, Hyunseul Jee, Kyung Chul Moon, Chaewon Kim, Minkyeong Choi, In Su Park, Chae Seung Lim
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Plasmodium</i> spp <i>Plasmodium falciparum</i> <i>Plasmodium vivax</i> LAMP multiplex LAMP Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/5e222a96d85e4de79e3420170944990d
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spelling	oai:doaj.org-article:5e222a96d85e4de79e3420170944990d2021-11-25T17:20:09ZDevelopment of a Multiplex Loop-Mediated Isothermal Amplification Assay for Diagnosis of <i>Plasmodium</i> spp., <i>Plasmodium falciparum</i> and <i>Plasmodium vivax</i>10.3390/diagnostics111119502075-4418https://doaj.org/article/5e222a96d85e4de79e3420170944990d2021-10-01T00:00:00Zhttps://www.mdpi.com/2075-4418/11/11/1950https://doaj.org/toc/2075-4418Malaria, caused by the parasite <i>Plasmodium</i> and transmitted by mosquitoes, is an epidemic that mainly occurs in tropical and subtropical regions. As treatments differ across species of malarial parasites, there is a need to develop rapid diagnostic methods to differentiate malarial species. Herein, we developed a multiplex malaria Pan/Pf/Pv/actin beta loop-mediated isothermal amplification (LAMP) to diagnose <i>Plasmodium</i> spp., <i>P. falciparum</i>, and <i>P. vivax</i>, as well as the internal control (IC), within 40 min. The detection limits of the multiplex malaria Pan/Pf/Pv/IC LAMP were 1 × 10<sup>2</sup>, 1 × 10<sup>2</sup>, 1 × 10<sup>2</sup>, and 1 × 10<sup>3</sup> copies/µL for four vectors, including the 18S rRNA gene (<i>Plasmodium</i> spp.), lactate dehydrogenase gene (<i>P. falciparum</i>), 16S rRNA gene (<i>P. vivax</i>), and human actin beta gene (IC), respectively. The performance of the LAMP assay was compared and evaluated by evaluating 208 clinical samples (118 positive and 90 negative samples) with the commercial RealStar<sup>®</sup> Malaria S&T PCR Kit 1.0. The developed multiplex malaria Pan/Pf/Pv/IC LAMP assay showed comparable sensitivity (100%) and specificity (100%) with the commercial RealStar<sup>®</sup> Malaria S&T PCR Kit 1.0 (100%). These results suggest that the multiplex malaria Pan/Pf/Pv/IC LAMP could be used as a point-of-care molecular diagnostic test for malaria.Woong Sik JangDa Hye LimYoungLan ChoeHyunseul JeeKyung Chul MoonChaewon KimMinkyeong ChoiIn Su ParkChae Seung LimMDPI AGarticle<i>Plasmodium</i> spp<i>Plasmodium falciparum</i><i>Plasmodium vivax</i>LAMPmultiplex LAMPMedicine (General)R5-920ENDiagnostics, Vol 11, Iss 1950, p 1950 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Plasmodium</i> spp <i>Plasmodium falciparum</i> <i>Plasmodium vivax</i> LAMP multiplex LAMP Medicine (General) R5-920
spellingShingle	<i>Plasmodium</i> spp <i>Plasmodium falciparum</i> <i>Plasmodium vivax</i> LAMP multiplex LAMP Medicine (General) R5-920 Woong Sik Jang Da Hye Lim YoungLan Choe Hyunseul Jee Kyung Chul Moon Chaewon Kim Minkyeong Choi In Su Park Chae Seung Lim Development of a Multiplex Loop-Mediated Isothermal Amplification Assay for Diagnosis of <i>Plasmodium</i> spp., <i>Plasmodium falciparum</i> and <i>Plasmodium vivax</i>
description	Malaria, caused by the parasite <i>Plasmodium</i> and transmitted by mosquitoes, is an epidemic that mainly occurs in tropical and subtropical regions. As treatments differ across species of malarial parasites, there is a need to develop rapid diagnostic methods to differentiate malarial species. Herein, we developed a multiplex malaria Pan/Pf/Pv/actin beta loop-mediated isothermal amplification (LAMP) to diagnose <i>Plasmodium</i> spp., <i>P. falciparum</i>, and <i>P. vivax</i>, as well as the internal control (IC), within 40 min. The detection limits of the multiplex malaria Pan/Pf/Pv/IC LAMP were 1 × 10<sup>2</sup>, 1 × 10<sup>2</sup>, 1 × 10<sup>2</sup>, and 1 × 10<sup>3</sup> copies/µL for four vectors, including the 18S rRNA gene (<i>Plasmodium</i> spp.), lactate dehydrogenase gene (<i>P. falciparum</i>), 16S rRNA gene (<i>P. vivax</i>), and human actin beta gene (IC), respectively. The performance of the LAMP assay was compared and evaluated by evaluating 208 clinical samples (118 positive and 90 negative samples) with the commercial RealStar<sup>®</sup> Malaria S&T PCR Kit 1.0. The developed multiplex malaria Pan/Pf/Pv/IC LAMP assay showed comparable sensitivity (100%) and specificity (100%) with the commercial RealStar<sup>®</sup> Malaria S&T PCR Kit 1.0 (100%). These results suggest that the multiplex malaria Pan/Pf/Pv/IC LAMP could be used as a point-of-care molecular diagnostic test for malaria.
format	article
author	Woong Sik Jang Da Hye Lim YoungLan Choe Hyunseul Jee Kyung Chul Moon Chaewon Kim Minkyeong Choi In Su Park Chae Seung Lim
author_facet	Woong Sik Jang Da Hye Lim YoungLan Choe Hyunseul Jee Kyung Chul Moon Chaewon Kim Minkyeong Choi In Su Park Chae Seung Lim
author_sort	Woong Sik Jang
title	Development of a Multiplex Loop-Mediated Isothermal Amplification Assay for Diagnosis of <i>Plasmodium</i> spp., <i>Plasmodium falciparum</i> and <i>Plasmodium vivax</i>
title_short	Development of a Multiplex Loop-Mediated Isothermal Amplification Assay for Diagnosis of <i>Plasmodium</i> spp., <i>Plasmodium falciparum</i> and <i>Plasmodium vivax</i>
title_full	Development of a Multiplex Loop-Mediated Isothermal Amplification Assay for Diagnosis of <i>Plasmodium</i> spp., <i>Plasmodium falciparum</i> and <i>Plasmodium vivax</i>
title_fullStr	Development of a Multiplex Loop-Mediated Isothermal Amplification Assay for Diagnosis of <i>Plasmodium</i> spp., <i>Plasmodium falciparum</i> and <i>Plasmodium vivax</i>
title_full_unstemmed	Development of a Multiplex Loop-Mediated Isothermal Amplification Assay for Diagnosis of <i>Plasmodium</i> spp., <i>Plasmodium falciparum</i> and <i>Plasmodium vivax</i>
title_sort	development of a multiplex loop-mediated isothermal amplification assay for diagnosis of <i>plasmodium</i> spp., <i>plasmodium falciparum</i> and <i>plasmodium vivax</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/5e222a96d85e4de79e3420170944990d
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Development of a Multiplex Loop-Mediated Isothermal Amplification Assay for Diagnosis of <i>Plasmodium</i> spp., <i>Plasmodium falciparum</i> and <i>Plasmodium vivax</i>

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