Principles and Applications of Loop-Mediated Isothermal Amplification to Point-of-Care Tests

Overview

Journal Biosensors (Basel)

Specialty Biotechnology

Date 2022 Oct 27

PMID 36290994

Authors

Jee-Woong Park

Affiliations

Soon will be listed here.

Abstract

For the identification of nucleic acids, which are important biomarkers of pathogen-mediated diseases and viruses, the gold standard for NA-based diagnostic applications is polymerase chain reaction (PCR). However, the requirements of PCR limit its application as a rapid point-of-care diagnostic technique. To address the challenges associated with regular PCR, many isothermal amplification methods have been developed to accurately detect NAs. Isothermal amplification methods enable NA amplification without changes in temperature with simple devices, as well as faster amplification times compared with regular PCR. Of the isothermal amplifications, loop-mediated isothermal amplification (LAMP) is the most studied because it amplifies NAs rapidly and specifically. This review describes the principles of LAMP, the methods used to monitor the process of LAMP, and examples of biosensors that detect the amplicons of LAMP. In addition, current trends in the application of LAMP to smartphones and self-diagnosis systems for point-of-care tests are also discussed.

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References

Kampeera J, Pasakon P, Karuwan C, Arunrut N, Sappat A, Sirithammajak S . Point-of-care rapid detection of Vibrio parahaemolyticus in seafood using loop-mediated isothermal amplification and graphene-based screen-printed electrochemical sensor. Biosens Bioelectron. 2019; 132:271-278. DOI: 10.1016/j.bios.2019.02.060. View

Das D, Lin C, Kwon J, Chuang H . Rotational diffusometric sensor with isothermal amplification for ultra-sensitive and rapid detection of SARS-CoV-2 nsp2 cDNA. Biosens Bioelectron. 2022; 210:114293. PMC: 9020650. DOI: 10.1016/j.bios.2022.114293. View

Martin A, Bouffier L, Grant K, Limoges B, Marchal D . Real-time electrochemical LAMP: a rational comparative study of different DNA intercalating and non-intercalating redox probes. Analyst. 2016; 141(13):4196-203. DOI: 10.1039/c6an00867d. View

Tanner N, Evans Jr T . Loop-mediated isothermal amplification for detection of nucleic acids. Curr Protoc Mol Biol. 2014; 105:Unit 15.14.. DOI: 10.1002/0471142727.mb1514s105. View

Compton J . Nucleic acid sequence-based amplification. Nature. 1991; 350(6313):91-2. DOI: 10.1038/350091a0. View

Goto M, Honda E, Ogura A, Nomoto A, Hanaki K . Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue. Biotechniques. 2009; 46(3):167-72. DOI: 10.2144/000113072. View

Kaarj K, Akarapipad P, Yoon J . Simpler, Faster, and Sensitive Zika Virus Assay Using Smartphone Detection of Loop-mediated Isothermal Amplification on Paper Microfluidic Chips. Sci Rep. 2018; 8(1):12438. PMC: 6102244. DOI: 10.1038/s41598-018-30797-9. View

Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N . Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 2000; 28(12):E63. PMC: 102748. DOI: 10.1093/nar/28.12.e63. View

Fischbach J, Xander N, Frohme M, Glokler J . Shining a light on LAMP assays--a comparison of LAMP visualization methods including the novel use of berberine. Biotechniques. 2015; 58(4):189-94. DOI: 10.2144/000114275. View

10.

Nzelu C, Gomez E, Caceres A, Sakurai T, Martini-Robles L, Uezato H . Development of a loop-mediated isothermal amplification method for rapid mass-screening of sand flies for Leishmania infection. Acta Trop. 2014; 132:1-6. DOI: 10.1016/j.actatropica.2013.12.016. View

11.

Diego J, Fernandez-Soto P, Marquez-Sanchez S, Santos Santos D, Febrer-Sendra B, Crego-Vicente B . SMART-LAMP: A Smartphone-Operated Handheld Device for Real-Time Colorimetric Point-of-Care Diagnosis of Infectious Diseases via Loop-Mediated Isothermal Amplification. Biosensors (Basel). 2022; 12(6). PMC: 9221248. DOI: 10.3390/bios12060424. View

12.

Takayama I, Nakauchi M, Takahashi H, Oba K, Semba S, Kaida A . Development of real-time fluorescent reverse transcription loop-mediated isothermal amplification assay with quenching primer for influenza virus and respiratory syncytial virus. J Virol Methods. 2019; 267:53-58. PMC: 7113748. DOI: 10.1016/j.jviromet.2019.02.010. View

13.

Lucchi N, Ljolje D, Silva-Flannery L, Udhayakumar V . Use of Malachite Green-Loop Mediated Isothermal Amplification for Detection of Plasmodium spp. Parasites. PLoS One. 2016; 11(3):e0151437. PMC: 4788150. DOI: 10.1371/journal.pone.0151437. View

14.

Rodriguez-Manzano J, Karymov M, Begolo S, Selck D, Zhukov D, Jue E . Reading Out Single-Molecule Digital RNA and DNA Isothermal Amplification in Nanoliter Volumes with Unmodified Camera Phones. ACS Nano. 2016; 10(3):3102-13. PMC: 4819493. DOI: 10.1021/acsnano.5b07338. View

15.

Lin M, Li Z, Lin Q, Wang P, Liu W, Yuan J . Development and clinical application of a rapid and visual loop-mediated isothermal amplification test for tetM gene in Clostridioides difficile strains cultured from feces. Int J Infect Dis. 2022; 122:676-684. DOI: 10.1016/j.ijid.2022.07.032. View

16.

Stedtfeld R, Tourlousse D, Seyrig G, Stedtfeld T, Kronlein M, Price S . Gene-Z: a device for point of care genetic testing using a smartphone. Lab Chip. 2012; 12(8):1454-62. DOI: 10.1039/c2lc21226a. View

17.

Wu G, Chen S, Levin R . Application of ethidium bromide monoazide for quantification of viable and dead cells of Salmonella enterica by real-time loop-mediated isothermal amplification. J Microbiol Methods. 2015; 117:41-8. DOI: 10.1016/j.mimet.2015.07.012. View

18.

Johnson G, Zubrzycki A, Henry M, Ranadheera C, Corbett C, Meyers A . Clinical evaluation of the GeneXpert® Xpert® Xpress SARS-CoV-2/Flu/RSV combination test. J Clin Virol Plus. 2022; 1(1):100014. PMC: 8040320. DOI: 10.1016/j.jcvp.2021.100014. View

19.

Choopara I, Arunrut N, Kiatpathomchai W, Dean D, Somboonna N . Rapid and visual Chlamydia trachomatis detection using loop-mediated isothermal amplification and hydroxynaphthol blue. Lett Appl Microbiol. 2016; 64(1):51-56. DOI: 10.1111/lam.12675. View

20.

Nguyen H, Bui H, Phan V, Seo T . An internet of things-based point-of-care device for direct reverse-transcription-loop mediated isothermal amplification to identify SARS-CoV-2. Biosens Bioelectron. 2021; 195:113655. PMC: 8458107. DOI: 10.1016/j.bios.2021.113655. View