Loop-mediated Isothermal Amplification LAMP, Simple Alternative Technique of Molecular Diagnosis Process in Medicals Analysis: A Review

Main Article Content

T. Uwiringiyeyezu
B. El Khalfi
J. Belhachmi
A. Soukri


The Medical diagnostic process requires more various methods to cover all sorts of clinical demands. Those methods depend on the types of samples and the requested analysis. The Medical diagnostic is considered as a process until making a clinical decision about a physiopathology or infection through medicals analysis. Nucleic acid amplification methods continue to play a role in these medicals analysis processes, but, many gold conventional standard Methods are time-consumers, less sensitive, technically challenging. It is that reason why recently, after the development of the PCR technique; the real-time PCR became a frequently requested tool to diagnose the virus, bacteria, fungal and parasites infections in different pathologies, especially in developed countries. However, its availability remains a big problem in resource-limited countries like Africa continent. It is because of the issue due to cost, technology, human resource constraints and the remote areas with limited access to laboratory facilities which stays also a major problem.

Nucleic acids amplification methods remain to be expensive, not accessible for all laboratories. In addition to this, they are easily inhibited and contaminated by the other biochemical reagents time-consumer, for this reason, other methods of nucleic acid amplification under isothermal conditions have emerged each with its  particularities. Despite their successes, molecular biology should have a method that combines many of the advantages and accessible everywhere. So, in 2000, the LAMP method was developed and offered many advantages such as speed, specificity, high sensitivity and the cost of very low equipment. In this review, we present the different studies to confirm the advantages of LAMP that we consider as an alternative to PCR in molecular  diagnostics with a wide range of use on clinical samples and in scientific research and also, the method to save the Africans patients from the high cost and time consuming of other amplification methods.

Polymerase chain reaction, loop-mediated isothermal amplification, molecular diagnosis, biological samples, diagnostic methods, medicals analysis

Article Details

How to Cite
Uwiringiyeyezu, T., Khalfi, B. E., Belhachmi, J., & Soukri, A. (2019). Loop-mediated Isothermal Amplification LAMP, Simple Alternative Technique of Molecular Diagnosis Process in Medicals Analysis: A Review. Annual Research & Review in Biology, 33(1), 1-12. https://doi.org/10.9734/arrb/2019/v33i130109
Review Article


Reddy AK, Balne PK, Reddy RK, Mathai A, Kaur I. Development and evaluation of loop-mediated isothermal amplification assay for rapid and inexpensive detection of cytomegalovirus DNA in vitreous specimens from suspected cases of viral retinitis. Journal of Clinical Microbiology. 2010;48(6):2050-2052.

Birmpa A, Kalogeropoulos K, Kokkinos P, Vantarakis A. Evaluation of two loop-mediated isothermal amplification methods for the detection of Salmonella enterica and Listeria monocytogenes in artificially contaminated ready-to-eat fresh produce. Italian Journal of Food Safety. 2015;4(3).

Tatibana BT, Sano A, Uno J, Kamei K, Igarashi T, Itano EN, et al. Detection of Paracoccidioides brasiliensis gp43 Gene in Sputa by Loop‐Mediated Isothermal Amplification Method. Journal of Clinical Laboratory Analysis. 2009;23(2):139-143.

Njiru ZK, Mikosza ASJ, Matovu E, Enyaru JCK, Ouma JO, Ndung’u JM, et al. African trypanosomiasis: Sensitive and rapid detection of the sub-genus Trypanozoon by loop-mediated isothermal amplification (LAMP) of parasite DNA. International Journal for Parasitology. 2008;38(5):589-599.

Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, et al. Loop-mediated isothermal amplification of DNA. Nucleic Acids Research. 2000;28 (12):e63-e63.

Knox CM, Chandler D, Short GA, Margolis, TP. Polymerase chain reaction-based assays of vitreous samples for the diagnosis of viral retinitis: Use in diagnostic dilemmas. Ophthalmology. 1998;105(1): 37-45.

Madhavan HN, Priya K, Biswas J. Current perspectives of herpesviral retinitis and choroiditis. Indian Journal of Pathology & Microbiology. 2004;47(4):453-468.

Bakhtiari S, Alvandi A, Pajavand H, Navabi J, Najafi F, Abiri R. Development and diagnostic evaluation of loop-mediated isothermal amplification using a new gene target for rapid detection of Helicobacter pylori. Jundishapur Journal of Microbiology. 2016;9(5).

Bista BR, Ishwad C, Wadowsky RM, Manna P, Randhawa PS, Gupta G. Development of a loop-mediated isothermal amplification assay for rapid detection of BK virus. Journal of Clinical Microbiology. 2007;45(5):1581-1587.

Suzuki R, Yoshikawa T, Ihira M, Enomoto Y, Inagaki S, Matsumoto K. Development of the loop-mediated isothermal amplification method for rapid detection of cytomegalovirus DNA. Journal of Virological Methods. 2006;132(1-2):216-221.

Wang X, Li X, Hu S, Qu H, Zhang Y, Wang X, et al. Rapid detection of active human cytomegalovirus infection in pregnancy using loop-mediated isothermal amplification. Molecular Medicine Reports. 2015;12(2):2269-2274.

de Vries JJ, van der Eijk AA, Wolthers KC, Rusman LG, Pas SD, Vossen AC, et al. Real-time PCR versus viral culture on urine as a gold standard in the diagnosis of congenital cytomegalovirus infection. Journal of Clinical Virology. 2012;53(2): 167-170.

Yamazaki W, Taguchi M, Kawai T, Kawatsu K, Sakata J, Inoue K, et al. Comparison of loop-mediated isothermal amplification assay and conventional culture methods for detection of Campylobacter jejuni and Campylobacter coli in naturally contaminated chicken meat samples. Appl. Environ. Microbiol. 2009; 75(6):1597-1603.

Soheily Z, Soleimani M, Majidzadeh-Ardebili K. Detection of Mycoplasma contamination of cell culture by A loop-mediated isothermal amplification method. Cell Journal (Yakhteh). 2019;12(1).

Kim CK, Cho EA, Shin DM, Choi SW, Shin SY. Comparative evaluation of the loop-mediated isothermal amplification assay for detecting pulmonary tuberculosis. Annals of Laboratory Medicine. 2018; 38(2):119-124.

Wang G, Shang Y, Wang Y, Tian H, Liu X. Comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time PCR. Virology Journal. 2013; 10(1):138.

Wang X, Seo DJ, Lee MH, Choi C. Comparison of conventional PCR, multiplex PCR, and loop-mediated isothermal amplification assays for rapid detection of Arcobacter species. Journal of Clinical Microbiology. 2014;52(2):557-563.

Ni XW, MC Manus DP, Lou ZZ, Yang JF, Yan HB, Fan YL, et al. A comparison of Loop-Mediated Isothermal Amplification (LAMP) with other surveillance tools for Echinococcus granulosus diagnosis in canine definitive hosts. PLoS One. 2014; 9(7):e100877.

Sotiriadou I, Karanis P. Evaluation of loop-mediated isothermal amplification for detection of Toxoplasma gondii in water samples and comparative findings by polymerase chain reaction and immunofluorescence test (IFT). Diagnostic Microbiology and Infectious Disease. 2008; 62(4):357-365.

Anklam K, Kulow M, Yamazaki W, Döpfer D. Development of real-time PCR and Loop-Mediated Isothermal Amplification (LAMP) assays for the differential detection of digital dermatitis associated treponemes. PloS One. 2017;12(5): e0178349.

Ni XW, McManus DP, Lou ZZ, Yang JF, Yan HB, Li L, Fan YL .A comparison of Loop-Mediated Isothermal Amplification (LAMP) with other surveillance tools for Echinococcus granulosus diagnosis in canine definitive hosts. PLoS One. 2014; 9(7):e100877.

Bojang AL, Mendy FS, Tientcheu LD, Otu J, Antonio M, Kampmann B, Sutherland JS. Comparison of TB-LAMP, GeneXpert MTB/RIF and culture for diagnosis of pulmonary tuberculosis in The Gambia. Journal of Infection. 2016;72(3):332-337.

Misawa Y, Yoshida A, Saito R, Yoshida H, Okuzumi K, Ito N, Koike K. Application of loop-mediated isothermal amplification technique to rapid and direct detection of methicillin-resistant Staphylococcus aureus (MRSA) in blood cultures. Journal of Infection and Chemotherapy. 2007;13(3): 134-140.

Sema M, Alemu A, Bayih AG, Getie S, Getnet G, Guelig D, Pillai DR. Evaluation of non-instrumented nucleic acid amplification by loop-mediated isothermal amplification (NINA-LAMP) for the diagnosis of malaria in Northwest Ethiopia. Malaria Journal. 2015;14(1):44.

Kargar M, Askari A, Doosti A, Ghorbani-Dalini S. Loop-mediated isothermal amplification assay for rapid detection of hepatitis C virus. Indian Journal of Virology. 2012;23(1):18-23.

Bartolone SN, Tree MO, Conway MJ, Chancellor MB, Lamb LE. Reverse Transcription-Loop-Mediated Isothermal Amplification (RT-LAMP) assay for Zika virus and housekeeping genes in urine, serum and mosquito samples. JoVE (Journal of Visualized Experiments). 2018; (139):e58436.

Iwamoto T, Sonobe T, Hayashi K. Loop-mediated isothermal amplification for direct detection of Mycobacterium tuberculosis complex, Mycobacterium avium and M. intracellulare in sputum samples. Journal of Clinical Microbiology. 2003;41(6):2616-2622.

Yari F, Abiri R, Aryan E, Ahmadi Jouybari T, Navabi J, Alvandi A. Loop‐mediated isothermal amplification as a fast noninvasive method of helicobacter pylori diagnosis. Journal of Clinical Laboratory Analysis. 2016;30(5):464-470.

Poon LL, Leung CS, Chan KH, Lee JH, Yuen KY, Guan Y, et al. Detection of human influenza A viruses by loop-mediated isothermal amplification. Journal of Clinical Microbiology. 2005;43(1):427-430.

Noguchi H, Iwase T, Omagari D, Asano M, Nakamura R, Ueki K, et al. Rapid detection of Candida albicans in oral exfoliative cytology samples by loop-mediated isothermal amplification. Journal of Oral Science. 2017;59(4):541-547.

Niessen L, Vogel RF. Detection of Fusarium graminearum DNA using a Loop-Mediated Isothermal Amplification (LAMP) assay. International Journal of Food Microbiology. 2010;140(2-3):183-191.

Amirabadi AR, Shahhosseiny MH, Yousefi JV, Ghahri M. Loop Mediated Isothermal Amplification (LAMP) in diagnosis of neorocryptococcosis. African Journal of Biotechnology. 2012;11(17): 3986-3992.

Yo A, Yamamoto M, Nakayama T, Ishikawa J, Makimura K. Detection and identification of Trichophyton tonsurans from clinical isolates and hairbrush samples by loop‐mediated isothermal amplification system. The Journal of Dermatology. 2016;43(9):1037-1043.

Scheel CM, Zhou Y, Theodoro RC, Abrams B, Balajee SA, Litvintseva AP. Development of a loop-mediated isothermal amplification method for detection of Histoplasma capsulatum DNA in clinical samples. Journal of clinical Microbiology. 2014;52(2):483-488.

Tang Q, Tian S, Yu N, Zhang X, Jia X, Han L, et al. Development and evaluation of a loop-mediated isothermal amplification method for rapid detection of Aspergillus fumigatus. Journal of Clinical Microbiology. 2016;54(4):950-955.

Carrillo C, Werbajh S, Malnero C, Stolowicz F, Larocca L, Malirat V, et al. Development of a colorimetric RT-LAMP amplification assay adapted to an early and easy detection of Dengue virus. International Journal of Infectious Diseases. 2018;73:171.

Maryam S, Rasheed T, Latif A, Zahra R, Zahur AB, Farooq U, et al. One-step real-time loop-mediated isothermal amplification (RT-LAMP): Evaluation and its application for the detection of foot-and-mouth-disease virus and its serotypes. Turkish Journal of Veterinary and Animal Sciences. 2017;41(3):435-443.

Nie X. Reverse transcription loop-mediated isothermal amplification of DNA for detection of Potato virus Y. Plant Disease. 2005;89(6):605-610.

Su Y, Yang Y, Peng Q, Zhou D, Chen Y, Wang Z, et al. Development and application of a rapid and visual loop-mediated isothermal amplification for the detection of Sporisorium scitamineum in sugarcane. Scientific Reports. 2016;6: 23994.

Coelho B, Veigas B, Águas H, Fortunato E, Martins R, Baptista P, et al. A digital microfluidics platform for loop-mediated isothermal amplification detection. Sensors. 2016,17(11).

Wang Y, Wang Y, Lan R, Xu H, Ma A, Li D, et al. Multiple endonuclease restriction real-time loop-mediated isothermal amplification: A novel analytically rapid, sensitive, multiplex loop-mediated isothermal amplification detection technique. The Journal of Molecular Diagnostics. 2015;17(4):392-401.

Biswal D. Advances in Loop-Mediated Isothermal Amplification (LAMP) technology and its necessity to detect helminth infections: An overview. Biomarkers J. 2016;2(2):1-5.

De Koninck AS, Cnops L, Hofmans M, Jacobs J, Van den Bossche D, Philippé J. Diagnostic performance of the loop-mediated isothermal amplification (LAMP) based illumigene® malaria assay in a non-endemic region. Malaria Journal. 2017; 16(1):418.

Abbasi I, Kirstein OD, Hailu A, Warburg A. Optimization of Loop-Mediated Isothermal Amplification (LAMP) assays for the detection of Leishmania DNA in human blood samples. Acta Tropica. 2016;162: 20-26.

Seok Y, Joung HA, Byun JY, Jeon HS, Shin SJ, Kim S, et al. A paper-based device for performing loop-mediated isothermal amplification with real-time simultaneous detection of multiple DNA targets. Theranostics. 2017;7(8):2220.

Mikita K, Maeda T, Yoshikawa S, Ono T, Miyahira Y, Kawana A. The direct boil-Lamp method: A simple and rapid diagnostic method for cutaneous leishmaniasis. Parasitology International. 2014;63(6):785-789.

Nzelu CO, Cáceres AG, Guerrero-Quincho S, Tineo-Villafuerte E, Rodriquez-Delfin L, Hashiguchi Y, et al. A rapid molecular diagnosis of cutaneous leishmaniasis by colorimetric malachite green-Loop-Mediated Isothermal Amplification (LAMP) combined with an FTA card as a direct sampling tool. Acta Tropica. 2016;153: 116-119.

Du WF, Ge JH, Li JJ, Tang LJ, Yu RQ, Jiang JH. Single-step, high-specificity detection of single nucleotide mutation by primer-activatable loop-mediated isothermal amplification (PA-LAMP). Analytica chimica acta. 2019;1050:132-138.

Zhang T, Lu H, Chen C, Jiang H, Wu S. Genetic diversity of mtDNA D-loop and maternal origin of three Chinese native horse breeds. Asian-Australasian Journal of Animal Sciences. 2012;25(7):921.

Kumari S, Kumar RR, Mendiratta SK, Kumar D, Rana PD, Jawla J, et al. Species-specific Loop-Mediated Isothermal Amplification (LAMP) assay for identification of tissue of cattle origin by targeting mitochondrial gene sequences. 3 Biotech. 2019;9(3):69.

Gao X, Sun B, Guan Y. Pullulan reduces the non-specific amplification of Loop-Mediated Isothermal Amplification (LAMP). Analytical and Bioanalytical Chemistry. 2019;411(6):1211-1218.

Zhang J, Cao J, Zhu M, Xu M, Shi F. Loop-mediated isothermal amplification-lateral-flow dipstick (LAMP-LFD) to detect Mycoplasma ovipneumoniae. World Journal of Microbiology and Biotechnology. 2019;35(2):31.

Kikkawa HS, Aragane M, Tsuge K. Species identification of white false hellebore (Veratrum album subsp. oxysepalum) by Loop-Mediated Isothermal Amplification (LAMP). Forensic Toxicology. 2019;37(2):308-315.

Wan L, Gao J, Chen T, Dong C, Li H, Martins RP, et al. Lamp Port: A handheld digital microfluidic device for loop-mediated isothermal amplification (LAMP). Biomedical microdevices. 2019;21(1):9.

Kurosaki Y, Magassouba NF, Oloniniyi OK, Cherif MS, Sakabe S, Yasuda J, et al. Development and evaluation of reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay coupled with a portable device for rapid diagnosis of Ebola virus disease in Guinea. PLoS Neglected Tropical Diseases. 2016;10(2): e0004472.

Tian W, Li P, He W, Liu C, Li Z. Rolling circle extension-actuated Loop-Mediated Isothermal Amplification (RCA-LAMP) for ultrasensitive detection of micro RNAs. Biosensors and Bioelectronics. 2019;128: 17-22.

Enomoto Y, Yoshikawa T, Ihira M, Akimoto S, Miyake F, Asano Y, et al. Rapid diagnosis of herpes simplex virus infection by a loop-mediated isothermal amplification method. Journal of Clinical Microbiology. 2005;43(2):951-955.

Rödel J, Bohnert JA, Stoll S, Wassill L, Edel B, Pfister W, et al. Evaluation of loop-mediated isothermal amplification for the rapid identification of bacteria and resistance determinants in positive blood cultures. European Journal of Clinical Microbiology & Infectious Diseases. 2017; 36(6):1033-1040.

Yeh HY, Shoemaker CA, Klesius PH. Evaluation of a loop-mediated isothermal amplification method for rapid detection of channel catfish Ictalurus punctatus important bacterial pathogen Edwardsiella ictaluri. Journal of microbiological methods, 2005;63(1):36-44.

Mamba TS, Mbae CK, Kinyua J, Mulinge E, Mburugu GN, Njiru ZK. Lateral flow loop-mediated isothermal amplification test with stem primers: Detection of cryptosporidium species in Kenyan children presenting with diarrhea. Journal of Tropical Medicine; 2018.

Tambo M, Auala JR, Sturrock HJ, Kleinschmidt I, Bock R, Mumbengegwi DR, et al. Evaluation of loop-mediated isothermal amplification as a surveillance tool for malaria in reactive case detection moving towards elimination. Malaria Journal. 2008;17(1):255.

Modak SS, Barber CA, Geva E, Abrams WR, Malamud D, Ongagna YSY. Rapid point-of-care isothermal amplification assay for the detection of malaria without nucleic acid purification. Infectious Diseases: Research and Treatment. 2016; 9.

Tomlinson J. In-field diagnostics using loop-mediated isothermal amplification. In Phytoplasma. 2013;291-300. Humana Press, Totowa, NJ.

Sirichaisinthop J, Buates S, Watanabe R, Han ET, Suktawonjaroenpon W, Krasaesub S, Sattabongkot J, et al. Evaluation of Loop-Mediated Isothermal Amplification (LAMP) for malaria diagnosis in a field setting. The American Journal of Tropical Medicine and Hygiene. 2011; 85(4):594-596.

Poon LL, Wong BW, Ma EH, Chan KH, Chow LM, Peiris JM, et al. Sensitive and inexpensive molecular test for falciparum malaria: detecting Plasmodium falciparum DNA directly from heat-treated blood by loop-media ted isothermal amplification. Clinical Chemistry. 2006;52(2):303-306.

Cook J, Aydin-Schmidt B, González IJ, Bell D, Edlund E, Bjork Man A, et al. Loop-Mediated Isothermal Amplification (LAMP) for point-of-care detection of asymptomatic low-density malaria parasite carriers in Zanzibar. Malaria Journal. 2015; 14(1):43.