CC BY-NC-ND 4.0 · Journal of Health and Allied Sciences NU 2017; 07(03): 042-048
DOI: 10.1055/s-0040-1708723
Review Article

Loop-mediated isothermal amplification (LAMP) : A rapid molecular diagnosis technique for detection of human pathogens

Gunimala Chakraborty
1  DST women scientist, Division of Infectious Diseases, Nitte University Centre for Science Education and Research Paneer Campus, Nitte University, Mangalore, Karnataka, India.
Indrani Karunasagar
2  Director (R&D), Dean, Faculty of Biological Sciences, Nitte University, Mangalore, Karnataka, India.
Anirban Chakraborty
3  Deputy Director, Nitte University Centre for Science Education and Research, Nitte University, Mangalore, Karnataka, India.
› Author Affiliations


Delivery of quality healthcare in case of an infectious disease depends on how efficiently and how quickly the responsible pathogens are detected from the samples. Molecular methods can detect the presence of pathogens in a rapid and sensitive manner. Over the years, a number of such assays have been developed. However, these methods, although highly reliable and efficient, require use of expensive equipment, reagents, and trained personnel. Therefore, development of molecular assays that are simple, rapid, cost-effective, yet sensitive, is highly warranted to ensure efficient management or treatment strategies. Loop-mediated isothermal amplification (LAMP), a technique invented in the year 2000, is a novel method that amplifies DNA at isothermal conditions. Since its invention, this technique has been one of the most extensively used molecular diagnostic tools in the field of diagnostics offering rapid, accurate and cost-effective diagnosis of infectious diseases. Using the LAMP principle, many commercial kits have been developed in the last decade for a variety of human pathogens including bacteria, viruses and parasites. Currently LAMP assay is being considered as an effective diagnostic tool for use in developing countries because of its simple working protocol, allowing even an onsite application. The focus of this review is to describe the salient features of this technique the current status of development of LAMP assays with an emphasis on the pathogens of clinical significance.

Publication History

Received: 27 March 2017

Accepted: 23 August 2017

Publication Date:
22 April 2020 (online)

© .

Thieme Medical and Scientific Publishers Private Ltd.
A-12, Second Floor, Sector -2, NOIDA -201301, India

  • References

  • 1 Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N. Enzymatic amplification of b-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anaemia. Science. 1985 Dec 20; 230(4732):1350-4.
  • 2 Mullis K, Faloona F, Scharf S, Saiki RK, Horn GT, Erlich H. Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. InCold Spring Harbor symposia on quantitative biology 1986 Jan 1 (Vol. 51, pp. 263-273). Cold Spring Harbor Laboratory Press.
  • 3 Lauerman LH. Advances in PCR technology. Animal health research reviews. 2004 Dec 1; 5(02):247-8.
  • 4 Black EM, Lowings JP, Smith J, Heaton PR, McElhinney LM. A rapid RTPCR method to differentiate six established genotypes of rabies and rabies-related viruses using TaqMan™ technology. Journal of virological methods. 2002 Aug 31; 105(1):25-35.
  • 5 Deb R, Chakraborty S. Trends in veterinary diagnostics. Journal of Veterinary Science and Technology. 2012; 3(1).
  • 6 Dhama K, Mahendran M, Tomar S, Thomas P. New generation techniques for detection and characterization of viral pathogens of poultry. Poult. Fortune. 2008; 9(8):32-6.
  • 7 Riaz F, Rashid I,Akbar H, Shehzad W, Islam S, Bajwa AA, Ashraf KSaeed K,. DNA Amplification Techniques for the Detection of Toxoplasma gondii Tissue Cysts in Meat Producing Animals: A Narrative Review 1 Article.K Iran J Parasitol. 2016 Oct-Dec; 11(4):431-440.
  • 8 Singh SD, Barathidasan R, Kumar A, Deb R, Verma AK, Dhama K. Recent trends in diagnosis and control of Marek's Disease (MD) in poultry. Pakistan Journal of Biological Sciences. 2012 Oct 15; 15(20):964.
  • 9 Nath G, Maurya P, Gulati AK. ERIC PCR and RAPD based fingerprinting of Salmonella Typhi strains isolated over a period of two decades. Infection, genetics and evolution. 2010 May 31; 10(4):530-6.
  • 10 Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T. Loop-mediated isothermal amplification of DNA. Nucleic acids research. 2000 Jun 15; 28(12).
  • 11 .Fu S, Qu G, Guo S, Ma L, Zhang N, Zhang S, Gao S, Shen Z. Applications of loop-mediated isothermal DNA amplification. Applied biochemistry and biotechnology. 2011 Apr 1; 163(7):845-50.
  • 12 Nagamine K, Watanabe K, Ohtsuka K, Hase T, Notomi T. Loop-mediated isothermal amplification reaction using a nondenatured template. Clinical Chemistry. 2001 Sep 1; 47(9):1742-3.
  • 13 Nagamine K, Hase T, Notomi T. Accelerated reaction by loop-mediated isothermal amplification using loop primers. Molecular and cellular probes. 2002 Jun 30; 16(3):223-9.
  • 14 Nagamine K, Kuzuhara Y, Notomi T. Isolation of single-stranded DNA from loop-mediated isothermal amplification products. Biochemical and biophysical research communications. 2002 Feb 1; 290(4):1195-8.
  • 15 Parida M, Sannarangaiah S, Dash PK, Rao PV, and Morita K. Loop mediated isothermal amplification (LAMP): a new generation of innovative gene amplification technique; perspectives in clinical diagnosis of infectious diseases. Reviews in medical virology. 2008 Nov 1; 18(6):407-21.
  • 16 Tomita N, Mori Y, Kanda H, Notomi T. Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products. Nature protocols. 2008 May 1; 3(5):877-82.
  • 17 Mori Y, Notomi T. Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases. Journal of infection and chemotherapy. 2009 Jan 1; 15(2):62-9.
  • 18 Mori Y, Kanda H, Notomi T. Loop-mediated isothermal amplification (LAMP): recent progress in research and development. Journal of infection and chemotherapy. 2013 Jan 1; 19(3):404-11.
  • 19 Lu X, Mo ZY, Zhao HB, Yan H, Shi L. LAMP-based method for a rapid identification of Legionella spp. and Legionella pneumophila. Applied microbiology and biotechnology. 2011 Oct 1; 92(1):179.
  • 20 Xu Z, Li L, Chu J, Peters BM, Harris ML, Li B, Shi L, Shirtliff ME. Development and application of loop-mediated isothermal amplification assays on rapid detection of various types of staphylococci strains. Food Research International. 2012 Jul 31; 47(2):166-73.
  • 21 Sonthayanon P, Chierakul W, Wuthiekanun V, Thaipadungpanit J, Kalambaheti T, Boonsilp S, Amornchai P, Smythe LD, Limmathurotsakul D, Day NP, Peacock SJ. Accuracy of loop-mediated isothermal amplification for diagnosis of human leptospirosis in Thailand. The American journal of tropical medicine and hygiene. 2011 Apr 5; 84(4):614-20.
  • 22 Hua-Wei Chen 201523 Chen HW, Weissenberger G, Atkins E, Chao CC, Suputtamongkol Y, Ching WM. Highly sensitive loop-mediated isothermal amplification for the detection of Leptospira. International journal of bacteriology. 2015 Jan 27, 23(1).
  • 23 Koizumi N, Nakajima C, Harunari T, Tanikawa T, Tokiwa T, Uchimura E, Furuya T, Mingala CN, Villanueva MA, Ohnishi M, Suzuki Y. A new loopmediated isothermal amplification method for rapid, simple, and sensitive detection of Leptospira spp. in urine. Journal of clinical microbiology. 2012 Jun 1; 50(6):2072-4.
  • 24 Tang MJ, Zhou S, Zhang XY, Pu JH, Ge QL, Tang XJ, Gao YS. Rapid and sensitive detection of Listeria monocytogenes by loop-mediated isothermal amplification. Current microbiology. 2011 Dec 1; 63(6):511.
  • 25 Kurosaki Y, Sakuma T, Fukuma A, Fujinami Y, Kawamoto K, Kamo N, Makino SI, Yasuda J. A simple and sensitive method for detection of Bacillus anthracis by loop-mediated isothermal amplification. Journal of applied microbiology. 2009 Dec 1; 107(6):1947-56.
  • 26 Yamazaki W, Seto K, Taguchi M, Ishibashi M, Inoue K. Sensitive and rapid detection of cholera toxin-producing Vibrio cholerae using a loop-mediated isothermal amplification. BMC microbiology. 2008 Jun 12; 8(1):94.
  • 27 Boyanton BL, Sural P, Loomis CR, Pesta C, Gonzalez-Krellwitz L, Robinson-Dunn B, Riska P. Loop-mediated isothermal amplification compared to real-time PCR and enzyme immunoassay for toxigenic Clostridium difficile detection. Journal of clinical microbiology. 2012 Mar 1; 50(3):640-5.
  • 28 Inácio J, Flores O, Spencer-Martins I. Efficient identification of clinically relevant Candida yeast species by use of an assay combining panfungal loop-mediated isothermal DNA amplification with hybridization to species-specific oligonucleotide probes. Journal of clinical microbiology. 2008 Feb 1; 46(2):713-20.
  • 29 Reza Ranjbar, Davoud Afshar. Development of a loop –mediated isothermal amplification assay for rapid detection of Yersinia enterocolitica via targeting a conserved locus.Iran J Microbiol. 2015 Aug; 7(4):185-190.
  • 30 Song L, Li J, Hou S, Li X, Chen S. Establishment of loop-mediated isothermal amplification (LAMP) for rapid detection of Brucella spp. and application to milk and blood samples. Journal of microbiological methods. 2012 Sep 30; 90(3):292-7.
  • 31 Hill J, Beriwal S, Chandra I, Paul VK, Kapil A, Singh T, Wadowsky RM, Singh V, Goyal A, Jahnukainen T, Johnson JR. Loop-mediated isothermal amplification assay for rapid detection of common strains of Escherichia coli. Journal of Clinical Microbiology. 2008 Aug 1; 46(8):2800-4.
  • 32 Thekisoe OM, Kuboki N, Nambota A, Fujisaki K, Sugimoto C, Igarashi I, Yasuda J, Inoue N. Species-specific loop-mediated isothermal amplification (LAMP) for diagnosis of trypanosomosis. Acta tropica. 2007 Jun 30; 102(3):182-9.
  • 33 Plutzer J, Karanis P. Rapid identification of Giardia duodenalis by loopmediated isothermal amplification (LAMP) from faecal and environmental samples and comparative findings by PCR and realtime PCR methods. Parasitology research. 2009 May 1; 104(6):1527- 33.
  • 34 Fernández-Soto P, Arahuetes JG, Hernández AS, Abán JL, Santiago BV, Muro A. A loop-mediated isothermal amplification (LAMP) assay for early detection of Schistosoma mansoni in stool samples: a diagnostic approach in a murine model. PLoS Negl Trop Dis. 2014 Sep 4; 8(9):e3126.
  • 35 Najafabadi ZG, Oormazdi H, Akhlaghi L, Meamar AR, Nateghpour M, Farivar L, Razmjou E. Detection of Plasmodium vivax and Plasmodium falciparum DNA in human saliva and urine: Loop-mediated isothermal amplification for malaria diagnosis. Acta Tropica. 2014 Aug 31; 136:44- 9.
  • 36 Poon LL, Leung CS, Chan KH, Lee JH, Yuen KY, Guan Y, Peiris JS. Detection of human influenza A viruses by loop-mediated isothermal amplification. Journal of clinical microbiology. 2005 Jan 1; 43(1):427- 30
  • 37 Khan MG, Bhaskar KR, Salam MA, Akther T, Pluschke G, Mondal D. Diagnostic accuracy of loop-mediated isothermal amplification (LAMP) for detection of Leishmania DNA in buffy coat from visceral leishmaniasis patients. Parasites & vectors. 2012 Dec 3; 5(1):280.
  • 38 Yamazaki W. Sensitive and rapid detection of Campylobacter jejuni and Campylobacter coli using loop-mediated isothermal amplification. PCR Detection of Microbial Pathogens. 2013:267-77.
  • 39 Nozomu Hanaoka, Minenosuke Matsutani, Masaaki Satoh, Motohiko Ogawa, Mutsunori Shirai, Shuji Ando. Development of a Novel Loop- Mediated Isothermal Amplification (LAMP) Assay for the Detection of Rickettsia spp. Japanese Journal of Infectious Diseases Vol. 70 No. 1.
  • 40 Jayawardena S. Loop-Mediated Isothermal Amplification for Influenza A (H5N1) Virus-Volume 13, Number 6—June 2007-Emerging Infectious Disease journal-CDC.
  • 41 Hagiwara M, Sasaki H, Matsuo K, Honda M, Kawase M, Nakagawa H. Loop-mediated isothermal amplification method for detection of human papillomavirus type 6, 11, 16, and 18. Journal of medical virology. 2007 May 1; 79(5):605-15.
  • 42 .M M Parida,S, R SanthoshP K, DashN K, Tripathi V , Lakshmi,N, Mamidi,A, ShrivastvaN, GuptaP, Saxena J, Pradeep Babul P, V Lakshmana Rao, Kouichi Morita.Rapid and Real-Time Detection of Chikungunya Virus by Reverse Transcription Loop-Mediated Isothermal Amplification Assay.J. Clin. Microbiol. February 2007 vol. 45 no.2 351-357.
  • 43 Toriniwa H, Komiya T. Rapid detection and quantification of Japanese encephalitis virus by real-time reverse transcription loop-mediated isothermal amplification. Microbiology and immunology. 2006 May 1; 50(5):379-87.
  • 44 Parida MM, Santhosh SR, Dash PK, Tripathi NK, Saxena P, Ambuj S, Sahni AK, Rao PL, Morita K. Development and evaluation of reverse transcription-loop-mediated isothermal amplification assay for rapid and real-time detection of Japanese encephalitis virus. Journal of Clinical Microbiology. 2006 Nov 1; 44(11):4172-8.
  • 45 Teoh BT, Sam SS, Tan KK, Johari J, Danlami MB, Hooi PS, Md-Esa R, AbuBakar S. Detection of dengue viruses using reverse transcriptionloop- mediated isothermal amplification. BMC infectious diseases. 2013 Aug 21; 13(1):387.
  • 46 Parida M, Horioke K, Ishida H, Dash PK, Saxena P, Jana AM, Islam MA, Inoue S, Hosaka N, Morita K. Rapid detection and differentiation of dengue virus serotypes by a real-time reverse transcription-loopmediated isothermal amplification assay. Journal of clinical microbiology. 2005 Jun 1; 43(6):2895-903.
  • 47 Parida M, Posadas G, Inoue S, Hasebe F, Morita K. Real-time reverse transcription loop-mediated isothermal amplification for rapid detection of West Nile virus. Journal of clinical microbiology. 2004 Jan 1; 42(1):257-63.
  • 48 Shukla J, Saxena D, Rathinam S, Lalitha P, Joseph CR, Sharma S, Soni M, Rao PV, Parida M. Molecular detection and characterization of West Nile virus associated with multifocal retinitis in patients from southern India. International Journal of Infectious Diseases. 2012 Jan 31; 16(1):e53-9.
  • 49 Kubo, T., M. Agoh, Q. Le Mai, K. Fukushima and H. Nishimura et al., 2010. Development of a reverse transcription-loop-mediated isothermal amplification assay for detection of pandemic (H1N1) 2009 virus as a novel molecular method for diagnosis of pandemic influenza in resource-limited settings. J. Clin. Microbiol. 48: 728-735.
  • 50 Moslemi E, Shahhosseiny MH, Javadi G, Praivar K, Sattari TN, Amini HK. Loop mediated isothermal amplification (LAMP) for rapid detection of HBV in Iran. African Journal of Microbiology Research. 2009 Aug 31; 3(8):439-45.
  • 51 Leo L. M. Poon1, Cynthia S. W. Leung,Kwok H. Chan, Jack H C Lee, Kwok Y, Yi Guan, Joseph S M Peiris.Detection of Human Influenza A Viruses by Loop-Mediated Isothermal Amplification. J. Clin. Microbiol. January 2005.vol. 43 no. 1 427-430.
  • 52 .Abo H, Okamoto K, Anraku M, Otsuki N, Sakata M, Icenogle J, Zheng Q, Kurata T, Kase T, Komase K, Takeda M. Development of an improved RTLAMP assay for detection of currently circulating rubella viruses. Journal of virological methods. 2014 Oct 31; 207:73-7.
  • 53 Fukuda S, Takao S, Kuwayama M, Shimazu Y, Miyazaki K. Rapid detection of norovirus from fecal specimens by real-time reverse transcription-loop-mediated isothermal amplification assay. Journal of clinical microbiology. 2006 Apr 1; 44(4):1376-81.
  • 54 Xia J F, X F Yan, H. Yu, D. Qu and J.E. Long, 2011. Simple and rapid detection of human enterovirus 71 by reverse-transcription and loopmediated isothermal amplification: Cryopreservation affected the detection ability. Diagn. Microbiol. Infect. Dis., 71: 244-251.
  • 55 Dougbeh-Chris Nyan Kevin L. Swinson. A method for rapid detection and genotype identification of hepatitis C virus 1–6 by one-step reverse transcription loop-mediated isothermal amplification International,2016Volume 43,( 30–36).
  • 56 Nkouawa A, Sako Y, Li T, Chen X, Wandra T, Swastika IK, Nakao M, Yanagida T, Nakaya K, Qiu D, Ito A. Evaluation of a loop-mediated isothermal amplification method using fecal specimens for differential detection of Taenia species from humans. Journal of clinical microbiology. 2010 Sep 1; 48(9):3350-2.
  • 57 Fernández-Soto P, Sánchez-Hernández A, Gandasegui J, Santos CB, López-Abán J, Saugar JM, Rodríguez E, Vicente B, Muro A. Strong- LAMP: A LAMP Assay for Strongyloides spp. Detection in Stool and Urine Samples. Towards the Diagnosis of Human Strongyloidiasis Starting from a Rodent Model. PLoS Negl Trop Dis. 2016 Jul 14; 10(7):004836.
  • 58 Yee-Ling Lau, Meng-Yee Lai, Mun-Yik Fong, Jenarun Jelip, Rohela Mahmud Loop-Mediated Isothermal Amplification Assay for Identification of Five HumanPlasmodiumSpecies in Malaysia. Am J Trop Med Hyg, 2016 Feb 3; 94(2): 336–339.
  • 59 Lukinmaa S, Nakari U, Eklund M, Siitonen A. Application of molecular genetic methods in diagnostics and epidemiology of food-borne bacterial pathogens . APMIS (2004), 12, 908-929.