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      Detection of helminths by loop-mediated isothermal amplification assay: a review of updated technology and future outlook

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          Abstract

          Background

          Helminths are endemic in more than half of the world’s countries, raising serious public health concerns. Accurate diagnosis of helminth infection is crucial to control strategies. Traditional parasitological methods, serological tests and PCR-based assays are the major means of the diagnosis of helminth infection, but they are time-consuming and/or expensive, and sometimes provide inaccurate results. Loop mediated isothermal amplification (LAMP) assay, a sensitive, simple and rapid method was therefore developed for detection of helminths. This study aims to discuss the current status of application of LAMP on helminths detection and to make a comprehensive evaluation about this updated technology and its future outlook by comparing with several other diagnostic methods.

          Main body

          This review summarizes LAMP assay applied for helminth detection and helminthiasis surveillance. The basic principle of LAMP is introduced to help better understand its characteristics and each reported assay is assessed mainly based on its detection sensitivity, specificity and limitations, in comparison with other common diagnostic tests. Moreover, we discuss the limitations of the assays so as to clarify some potential ways of improvement.

          Conclusions

          Here, we summarize and discuss the advantages, disadvantages and promising future of LAMP in heliminth detection, which is expected to help update current knowledge and future perspectives of LAMP in highly sensitive and specific diagnosis and surveillance of helminthiasis and other parasitic diseases, and can contribute to the elimination of the diseases from endemic areas.

          Electronic supplementary material

          The online version of this article (10.1186/s40249-019-0530-z) contains supplementary material, which is available to authorized users.

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          Most cited references161

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          Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation.

          The loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method that uses only one type of enzyme. One of the characteristics of the LAMP method is its ability to synthesize extremely large amount of DNA. Accordingly, a large amount of by-product, pyrophosphate ion, is produced, yielding white precipitate of magnesium pyrophosphate in the reaction mixture. Judging the presence or absence of this white precipitate allows easy distinction of whether nucleic acid was amplified by the LAMP method. Since an increase in the turbidity of the reaction mixture according to the production of precipitate correlates with the amount of DNA synthesized, real-time monitoring of the LAMP reaction was achieved by real-time measurement of turbidity. Copyright 2001 Academic Press.
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            Genetic variants within the genus Echinococcus identified by mitochondrial DNA sequencing.

            The pattern of species and strain variation within the genus Echinococcus is complex and controversial. In an attempt to characterise objectively the various species and strains, the sequence of a region of the mitochondrial cytochrome c oxidase subunit I (CO1) gene was determined for 56 Echinococcus isolates. Eleven different genotypes were detected, including 7 within Echinococcus granulosus, and these were used to categorise the isolates. The 4 generally accepted Echinococcus species were clearly distinguishable using this approach. In addition, the consensus view of the strain pattern within E. granulosus, based on a variety of criteria of differentiation, was broadly upheld. Very little variation was detected within Echinococcus multilocularis. Remarkable intra-strain homogeneity was found at the DNA sequence level. This region of the rapidly evolving mitochondrial genome is useful as a marker of species and strain identity and as a preliminary indication of evolutionary divergence within the genus Echinococcus.
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              Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue.

              Loop-mediated isothermal amplification (LAMP), a novel gene amplification method, enables the synthesis of larger amounts of both DNA and a visible byproduct--namely, magnesium pyrophosphate--without thermal cycling. A positive reaction is indicated by the turbidity of the reaction solution or the color change after adding an intercalating dye to the reaction solution, but the use of such dyes has certain limitations. Hydroxy naphthol blue (HNB), a metal indicator for calcium and a colorimetric reagent for alkaline earth metal ions, was used for a new colorimetric assay of the LAMP reaction. Preaddition of 120 microM HNB to the LAMP reaction solution did not inhibit amplification efficiency. A positive reaction is indicated by a color change from violet to sky blue. The LAMP reaction with HNB could also be carried out in a 96-well microplate, and the reaction could be measured at 650 nm with a microplate reader. The colorimetric LAMP method using HNB would be helpful for high-throughput DNA and RNA detection.
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                Author and article information

                Contributors
                dengmh6@mail2.sysu.edu.cn
                zhongly23@mail2.sysu.edn.cn
                kamolnetr.oka@mahidol.ac.th
                yanin.lim@mahidol.ac.th
                lvzhiyue@mail.sysu.edu.cn
                Journal
                Infect Dis Poverty
                Infect Dis Poverty
                Infectious Diseases of Poverty
                BioMed Central (London )
                2095-5162
                2049-9957
                25 March 2019
                25 March 2019
                2019
                : 8
                : 20
                Affiliations
                [1 ]ISNI 0000 0001 2360 039X, GRID grid.12981.33, Zhongshan School of Medicine, , Sun Yat-sen University, ; Guangzhou, 510080 China
                [2 ]ISNI 0000 0004 0369 313X, GRID grid.419897.a, Key Laboratory of Tropical Disease Control (Sun Yat-sen University), Ministry of Education, ; Guangzhou, 510080 China
                [3 ]ISNI 0000 0004 1937 0490, GRID grid.10223.32, Faculty of Tropical Medicine, , Mahidol University, ; Bangkok, 10400 Thailand
                [4 ]Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, 510080 China
                [5 ]ISNI 0000 0001 2360 039X, GRID grid.12981.33, Fifth Affiliated Hospital, Sun Yat-sen University, ; Guangzhou, 519000 China
                Article
                530
                10.1186/s40249-019-0530-z
                6432754
                30905322
                414d97dc-2edc-432a-a145-388489e07476
                © The Author(s). 2019

                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.

                History
                : 5 September 2018
                : 7 March 2019
                Funding
                Funded by: the National Key Research and Development Program of China
                Award ID: 2016YFC1202003
                Categories
                Scoping Review
                Custom metadata
                © The Author(s) 2019

                loop-mediated isothermal amplification,helminth,point-of-care-test,epidemiological surveillance,field survey

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