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      Loop‐mediated isothermal amplification (LAMP): a versatile technique for detection of micro‐organisms

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          Summary

          Loop‐mediated isothermal amplification ( LAMP) amplifies DNA with high specificity, efficiency and rapidity under isothermal conditions by using a DNA polymerase with high displacement strand activity and a set of specifically designed primers to amplify targeted DNA strands. Following its first discovery by Notomi et al. ( 2000 Nucleic Acids Res 28: E63), LAMP was further developed over the years which involved the combination of this technique with other molecular approaches, such as reverse transcription and multiplex amplification for the detection of infectious diseases caused by micro‐organisms in humans, livestock and plants. In this review, available types of LAMP techniques will be discussed together with their applications in detection of various micro‐organisms. Up to date, there are varieties of LAMP detection methods available including colorimetric and fluorescent detection, real‐time monitoring using turbidity metre and detection using lateral flow device which will also be highlighted in this review. Apart from that, commercialization of LAMP technique had also been reported such as lyophilized form of LAMP reagents kit and LAMP primer sets for detection of pathogenic micro‐organisms. On top of that, advantages and limitations of this molecular detection method are also described together with its future potential as a diagnostic method for infectious disease.

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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): principle, features, and future prospects.

            Loop-mediated isothermal amplification (LAMP), a newly developed gene amplification method, combines rapidity, simplicity, and high specificity. Several tests have been developed based on this method, and simplicity is maintained throughout all steps, from extraction of nucleic acids to detection of amplification. In the LAMP reaction, samples are amplified at a fixed temperature through a repetition of two types of elongation reactions occurring at the loop regions: self-elongation of templates from the stem loop structure formed at the 3'-terminal and the binding and elongation of new primers to the loop region. The LAMP reaction has a wide range of possible applications, including point-of-care testing, genetic testing in resource-poor settings (such as in developing countries), and rapid testing of food products and environmental samples.
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              Is Open Access

              Isothermal Amplification Methods for the Detection of Nucleic Acids in Microfluidic Devices

              Diagnostic tools for biomolecular detection need to fulfill specific requirements in terms of sensitivity, selectivity and high-throughput in order to widen their applicability and to minimize the cost of the assay. The nucleic acid amplification is a key step in DNA detection assays. It contributes to improving the assay sensitivity by enabling the detection of a limited number of target molecules. The use of microfluidic devices to miniaturize amplification protocols reduces the required sample volume and the analysis times and offers new possibilities for the process automation and integration in one single device. The vast majority of miniaturized systems for nucleic acid analysis exploit the polymerase chain reaction (PCR) amplification method, which requires repeated cycles of three or two temperature-dependent steps during the amplification of the nucleic acid target sequence. In contrast, low temperature isothermal amplification methods have no need for thermal cycling thus requiring simplified microfluidic device features. Here, the use of miniaturized analysis systems using isothermal amplification reactions for the nucleic acid amplification will be discussed.
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                Author and article information

                Contributors
                cheehy@upm.edu.my
                Journal
                J Appl Microbiol
                J. Appl. Microbiol
                10.1111/(ISSN)1365-2672
                JAM
                Journal of Applied Microbiology
                John Wiley and Sons Inc. (Hoboken )
                1364-5072
                1365-2672
                12 February 2018
                March 2018
                : 124
                : 3 ( doiID: 10.1111/jam.2018.124.issue-3 )
                : 626-643
                Affiliations
                [ 1 ] Department of Medical Microbiology and Parasitology Faculty of Medicine and Health Sciences Universiti Putra Malaysia Serdang Malaysia
                [ 2 ] Department of Parasitology Faculty of Medicine Tropical Infectious Diseases Research and Education Centre (TIDREC) University of Malaya Kuala Lumpur Malaysia
                [ 3 ] Centre of Excellence for Food Safety Research (FOSREC) Faculty of Food Science and Technology Universiti Putra Malaysia Serdang Malaysia
                Author notes
                [*] [* ] Correspondence

                Hui‐Yee Chee, Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health Science, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia.

                E‐mail: cheehy@ 123456upm.edu.my

                Article
                JAM13647 1976
                10.1111/jam.13647
                7167136
                29165905
                de540ddb-ba0b-4d16-812e-c95f251f3680
                © 2017 The Society for Applied Microbiology

                This article is being made freely available through PubMed Central as part of the COVID-19 public health emergency response. It can be used for unrestricted research re-use and analysis in any form or by any means with acknowledgement of the original source, for the duration of the public health emergency.

                History
                : 26 May 2017
                : 09 November 2017
                : 13 November 2017
                Page count
                Figures: 2, Tables: 0, Pages: 18, Words: 13424
                Funding
                Funded by: Ministry of Higher Education Malaysia
                Award ID: UPM/700‐2/1/LRGS/5526400
                Funded by: Universiti Putra Malaysia , open-funder-registry 10.13039/501100004530;
                Categories
                Review Article
                Review Articles
                Custom metadata
                2.0
                March 2018
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.8.0 mode:remove_FC converted:15.04.2020

                Microbiology & Virology
                biotechnology,detection,diagnosis,diseases,rapid methods
                Microbiology & Virology
                biotechnology, detection, diagnosis, diseases, rapid methods

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