Volume 94, Issue 6
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



For differential detection of , , and , loop-mediated isothermal amplification (LAMP) assay targeting the cytochrome oxidase subunit 1 gene has been recently developed and shown to be sensitive, specific, and effective. However, to achieve differential identification, one specimen requires three reaction mixtures containing a primer set of each species separately, which is complex and time consuming and increases the risk of cross-contamination. In this study, we developed a simple differential identification of human species using multiplex LAMP (mLAMP) in combination with dot enzyme-linked immunosorbent assay (dot-ELISA). Forward inner primers of , , and labeled with fluorescein isothiocyanate (FITC), digoxigenin (DIG), and tetramethylrhodamine (TAMRA), respectively, and biotin-labeled backward inner primers were used in mLAMP. The mLAMP assay succeeded in specific amplification of each respective target gene in a single tube. Furthermore, the mLAMP product from each species was easily distinguished by dot-ELISA with an antibody specific for FITC, DIG, or TAMRA. The mLAMP assay in combination with dot-ELISA will make identification of human species simpler, easier, and more practical.


Article metrics loading...

Loading full text...

Full text loading...



  1. Garcia HH, Gonzalez AE, Evans CA, Gilman RH, Cysticercosis Working Group in Peru, , 2003. Taenia solium cysticercosis. Lancet 362: 547556.[Crossref]
  2. Ito A, Nakao M, Wandra T, Suroso T, Okamoto M, Yamasaki H, Sako Y, Nakaya K, , 2005. Taeniasis and cysticercosis in Asia and the Pacific: present state of knowledge and perspectives. Southeast Asian J Trop Med Public Health 36 (Suppl 4): S123S130.
  3. World Health Organization, 2010. First WHO Report on Neglected Tropical Diseases: Working to Overcome the Global Impact of Neglected Tropical Diseases. Geneva, Switzerland: World Health Organization, 97102.
  4. Winkler AS, , 2012. Neurocysticercosis in sub-Saharan Africa: a review of prevalence, clinical characteristics, diagnosis, and management. Pathog Glob Health 106: 261274.[Crossref]
  5. Li T, Craig PS, Ito A, Chen X, Qiu D, Qiu J, Sato MO, Wandra T, Bradshaw H, Li L, Yang Y, Wang Q, , 2006. Taeniasis/cysticercosis in a Tibetan population in Sichuan Province, China. Acta Trop 100: 223231.[Crossref]
  6. Ito A, Wandra T, Li T, Dekumyoy P, Nkouawa A, Okamoto M, Budke CM, , 2014. The present situation of human taeniases and cysticercosis in Asia. Recent Pat Antiinfect Drug Discov 9: 173185.[Crossref]
  7. Schantz PM, Moore AC, Munoz JL, Hartman BJ, Schaefer JA, Perasaud D, Sarti E, Wilson M, Flisser A, Aron AM, , 1992. Neurocysticercosis in an Orthodox Jewish community in New York City. N Engl J Med 327: 692695.[Crossref]
  8. Sorvillo F, Wilkins P, Shafir S, Eberhard M, , 2011. Public health implications of cysticercosis acquired in the United States. Emerg Infect Dis 17: 16.[Crossref]
  9. Yanagida T, Sako Y, Nakao M, Nakaya K, Ito A, , 2012. Taeniasis and cysticercosis due to Taenia solium in Japan. Parasit Vectors 5: 18.[Crossref]
  10. Murrell KD, Murrell KD, , 2005. Epidemiology of taeniosis and cysticercosis. , ed. WHO/FAO/OIE Guidelines for the Surveillance, Prevention and Control of Taeniosis/Cysticercosis. Paris, France: OIE, WHO, FAO, 2743.
  11. Flisser A, Craig PS, Ito A, Palmer SR, Soulsby L, Torgerson PR, Brown DWG, , 2011. Cysticercosis and taeniosis: Taenia solium, Taenia saginata, and Taenia asiatica . , eds. Oxford Textbook of Zoonoses, Biology, Clinical Practice and Public Health Control. Oxford, United Kingdom: Oxford University Press, 627644.
  12. Li T, Chen X, Yanagida T, Wang H, Long C, Sako Y, Okamoto M, Wu Y, Giraudoux P, Raoul F, Nkouawa A, Nakao M, Craig PS, Ito A, , 2013. Detection of human taeniases in Tibetan endemic areas, China. Parasitology 140: 16021607.[Crossref]
  13. Wandra T, Ito A, Swastika K, Dharmawan NS, Sako Y, Okamoto M, , 2013. Taeniases and cysticercosis in Indonesia: past and present situations. Parasitology 140: 16081616.[Crossref]
  14. Mori Y, Notomi T, , 2009. Loop-mediated isothermal amplification (LAMP), a rapid, accurate, and cost-effective diagnostic method for infectious diseases. J Infect Chemother 15: 6269.[Crossref]
  15. Mori Y, Kanda H, Notomi T, , 2013. Loop-mediated isothermal amplification (LAMP): recent progress in research and development. J Infect Chemother 19: 404411.[Crossref]
  16. Nkouawa A, Sako Y, Nakao M, Nakaya K, Ito A, , 2009. Loop-mediated isothermal amplification method for differentiation and rapid detection of Taenia species. J Clin Microbiol 47: 168174.[Crossref]
  17. Sako Y, Nkouawa A, Yanagida T, Ito A, Kolpashchikov DM, Gerasimova YV, , 2013. Chapter 9. Loop-mediated isothermal amplification method for a differential identification of human Taenia tapeworms. , eds. Nuclear Acid Detection, Methods and Protocols, Methods in Molecular Biology Vol. 1039. New York, NY: Humana Press, 109120.[Crossref]
  18. Nkouawa A, Sako Y, Li T, Chen X, Wandra T, Swastika IK, Nakao M, Yanagida T, Nakaya K, Qiu D, Ito A, , 2010. Evaluation of a loop-mediated isothermal amplification method using fecal specimens for differential detection of Taenia species from humans. J Clin Microbiol 48: 33503352.[Crossref]
  19. Nkouawa A, Sako Y, Li T, Chen X, Nakao M, Yanagida T, Okamoto M, Giraudoux P, Raoul F, Nakaya K, Xiao N, Qiu J, Qiu D, Craig PS, Ito A, , 2012. Loop-mediated isothermal amplification method for a differential identification of Taenia tapeworms from human: application to a field survey. Parasitol Int 61: 723725.[Crossref]
  20. Yamasaki H, Allan JC, Sato MO, Nakao M, Sako Y, Nakaya Y, Qiu D, Mamuti W, Craig PS, Ito A, , 2004. DNA differential diagnosis of taeniasis and cysticercosis by multiplex PCR. J Clin Microbiol 42: 548553.[Crossref]
  21. Nakaya K, Mamuti W, Xiao N, Sato MO, Wandra T, Nakao M, Sako Y, Yamasaki H, Ishikawa Y, Craig PS, Schantz PM, Ito A, , 2006. Usefulness of severe combined immunodeficiency (scid) and inbred mice for studies of cysticercosis and echinococcosis. Parasitol Int 55 (Suppl): S91S97.[Crossref]
  22. Iseki H, Alhassan A, Ohta N, Thekisoe OM, Yokoyama N, Inoue N, Nambota A, Yasuda J, Igarashi I, , 2007. Development of a multiplex loop-mediated isothermal amplification (mLAMP) method for the simultaneous detection of bovine Babesia parasites. J Microbiol Methods 71: 281287.[Crossref]
  23. Lee MF, Chen YH, Peng CF, , 2009. Evaluation of reverse transcription loop-mediated isothermal amplification in conjunction with ELISA-hybridization assay for molecular detection of Mycobacterium tuberculosis . J Microbiol Methods 76: 174180.[Crossref]
  24. Lee MF, Chen YH, Hsu HJ, Peng CF, , 2010. One-tube loop-mediated isothermal amplification combined with restriction endonuclease digestion and ELISA for colorimetric detection of resistance to isoniazid, ethambutol and streptomycin in Mycobacterium tuberculosis isolates. J Microbiol Methods 83: 5358.[Crossref]
  25. Mahony J, Chong S, Bulir D, Ruyter A, Mwawasi K, Waltho D, , 2013. Multiplex loop-mediated isothermal amplification (M-LAMP) assay for the detection of influenza A/H1, A/H3 and influenza B can provide a specimen-to-result diagnosis in 40 min with single genome copy sensitivity. J Clin Virol 58: 127131.[Crossref]
  26. Yamazaki W, Mioulet V, Murray L, Madi M, Haga T, Misawa N, Horii Y, King DP, , 2013. Development and evaluation of multiplex RT-LAMP assays for rapid and sensitive detection of foot-and-mouth disease virus. J Virol Methods 192: 1824.[Crossref]
  27. Njiru ZK, , 2011. Rapid and sensitive detection of human African trypanosomiasis by loop-mediated isothermal amplification combined with a lateral-flow dipstick. Diagn Microbiol Infect Dis 69: 205209.[Crossref]
  28. Kaewphinit T, Arunrut N, Kiatpathomchai W, Santiwatanakul S, Jaratsing P, Chansiri K, , 2013. Detection of Mycobacterium tuberculosis by using loop-mediated isothermal amplification combined with a lateral flow dipstick in clinical samples. BioMed Res Int 2013: 926230.[Crossref]
  29. Yongkiettrakul S, Jaroenram W, Arunrut N, Chareanchim W, Pannengpetch S, Suebsing R, Kiatpathomchai W, Pornthanakasem W, Yuthavong Y, Kongkasuriyachai D, , 2014. Application of loop-mediated isothermal amplification assay combined with lateral flow dipstick for detection of Plasmodium falciparum and Plasmodium vivax . Parasitol Int 63: 777784.[Crossref]
  30. Jung JH, Oh SJ, Kim YT, Kim SY, Kim WJ, Jung J, Seo TS, , 2015. Combination of multiplex reverse-transcription loop-mediated isothermal amplification with an immunochromatographic strip for subtyping influenza A virus. Anal Chim Acta 853: 541547.[Crossref]
  31. Nurul Najian AB, Engku Nur Syafirah EA, Ismail N, Mohamed M, Yean CY, , 2016. Development of multiplex loop mediated isothermal amplification (m-LAMP) label-based gold nanoparticles lateral flow dipstick biosensor for detection of pathogenic Leptospira . Anal Chim Acta 903: 142148.[Crossref]

Data & Media loading...

  • Received : 16 Nov 2015
  • Accepted : 01 Mar 2016

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error