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      Loop Mediated Isothermal Amplification: Principles and Applications in Plant Virology


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          In the last decades, the evolution of molecular diagnosis methods has generated different advanced tools, like loop-mediated isothermal amplification (LAMP). Currently, it is a well-established technique, applied in different fields, such as the medicine, agriculture, and food industries, owing to its simplicity, specificity, rapidity, and low-cost efforts. LAMP is a nucleic acid amplification under isothermal conditions, which is highly compatible with point-of-care (POC) analysis and has the potential to improve the diagnosis in plant protection. The great advantages of LAMP have led to several upgrades in order to implement the technique. In this review, the authors provide an overview reporting in detail the different LAMP steps, focusing on designing and main characteristics of the primer set, different methods of result visualization, evolution and different application fields, reporting in detail LAMP application in plant virology, and the main advantages of the use of this technique.

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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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            Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases

            Loop-mediated isothermal amplification (LAMP) is an established nucleic acid amplification method offering rapid, accurate, and cost-effective diagnosis of infectious diseases. This technology has been developed into commercially available detection kits for a variety of pathogens including bacteria and viruses. The current focus on LAMP methodology is as a diagnostic system to be employed in resource-limited laboratories in developing countries, where many fatal tropical diseases are endemic. The combination of LAMP and novel microfluidic technologies such as Lab-on-a-chip may facilitate the realization of genetic point-of-care testing systems to be used by both developed and developing countries in the near future. This review will describe the historical, current, and future developments of such technologies.
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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.

                Author and article information

                Plants (Basel)
                Plants (Basel)
                06 April 2020
                April 2020
                : 9
                : 4
                [1 ]Department of Agricultural, Food and Forest Sciences, University of Palermo, 90128 Palermo, Italy; andreagiovanni.caruso@ 123456unipa.it (A.G.C.); patrizia.bella@ 123456unipa.it (P.B.); livio.torta@ 123456unipa.it (L.T.); stassiraf@ 123456gmail.com (R.S.)
                [2 ]Department of Agricultural, Forestry and Food Sciences, University of Turin, 10095 Turin, Italy; slavica.matic@ 123456unito.it
                [3 ]Council for Agricultural Research and Economics, Research Center for Plant Protection and Certification, 00156 Rome, Italy; antonio.tiberini@ 123456crea.gov.it
                [4 ]Institute for Sustainable Plant Protection, National Research Council (IPSP-CNR), 10135 Turin, Italy
                Author notes
                [* ]Correspondence: stefano.panno@ 123456unipa.it (S.P.); salvatore.davino@ 123456unipa.it (S.D.); Tel.: +39-0912-389-6049 (S.P. & S.D.)

                These authors contributed equally to this work.

                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).


                loop-mediated isothermal amplification,lamp,bst dna polymerase,primers,real-time monitoring,virus,viroids,plant virology


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