Volume 82, Issue 5
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



We have developed two loop-mediated isothermal amplification (LAMP) assays for specific detection of and based on the 18S ribosomal RNA (rRNA) and the small nucleolar RNA (snoRNA) genes, respectively. The detection limit of the assays is 100 fg and 1 pg for and , respectively, with reactions conducted in 60 minutes. The two LAMP assays were used in detection of and infections in comparison with polymerase chain reaction (PCR) for DNA samples extracted from bugs collected from palm trees in Panama. Out of a total of 52 DNA samples from bugs 17 (33%) and 14 (27%) were -positive by LAMP and PCR, respectively, while, 7 (13%) and 4 (8%) were -positive by LAMP and PCR, respectively. Further evaluation of these LAMP assays is needed, especially with specimens collected from human patients as well as blood kept for transfusion purposes.


Article metrics loading...

The graphs shown below represent data from March 2017
Loading full text...

Full text loading...



  1. Pavia PX, Vallejo GA, Montilla M, Nicholls RS, Puerta CJ, , 2007. Detection of Trypanosoma cruzi and Trypanosoma rangeli infections in triatomine vectors by amplification of the histone H2A/SIRE and the sno-RNA-C11 genes. Rev Inst Med Trop Sao Paulo 49: 2330.[Crossref] [Google Scholar]
  2. Vallejo GA, Guhl F, Chiari E, Macedo AM, , 1999. Species specific detection of Trypanosoma cruzi and Trypanosoma rangeli in vector and mammalian hosts by polymerase chain reaction amplification of kinetoplast minicircle DNA. Acta Trop 72: 203212.[Crossref] [Google Scholar]
  3. Sanchez-Guillen MC, Barnabe C, Guegan JF, Tibayrenc M, Velasquez-Rojas M, Martinez-Munguia J, Salgado-Rosas H, Torres-Rasgado E, Rosas-Ramirez MI, Perez-Fuentes R, , 2002. High prevalence anti-Trypanosoma cruzi antibodies, among blood donors in the state of Puebla, a non-endemic area of Mexico. Mem Inst Oswaldo Cruz 97: 947952.[Crossref] [Google Scholar]
  4. de Moraes MH, Guarneri AA, Girardi FP, Rodriques JB, Eger I, Tyler KM, Steindel M, Grisard EC, , 2008. Different serological cross-reactivity of Trypanosoma rangeli forms in Trypanosoma cruzi infected patients sera. Parasit Vectors 1: 20.[Crossref] [Google Scholar]
  5. Morales L, Romero I, Diez H, Del Portillo P, Montilla M, Nicholls S, Puerta C, , 2002. Characterization of a candidate Trypanosoma rangeli small nucleolar RNA gene and its application in a PCR-based parasite detection. Exp Parasitol 102: 7280.[Crossref] [Google Scholar]
  6. Stevens JR, Brisse S, Maudlin I, Holmes PH, Miles MA, , 2004. Systematics of trypanosomes of medical and veterinary importance. , eds. The Trypanosomiases. Oxfordshire, UK: CABI Publishing, 123.[Crossref] [Google Scholar]
  7. Avila HA, Pereira JB, Thieman O, de Paiva E, Degrave W, Morel CM, Simpson L, , 1993. Detection of Trypanosoma cruzi in blood specimens of chronic Chagas patients by polymerase chain reaction amplification of kinetoplast minicircle DNA: comparison with serology and xenodiagnosis. J Clin Microbiol 31: 24212426. [Google Scholar]
  8. Kuboki N, Inoue N, Sakurai T, Di Cello F, Grab DJ, Suzuki H, Sugimoto C, Igarashi I, , 2003. Loop-mediated isothermal amplification (LAMP) for detection of African trypanosomes. J Clin Microbiol 38: 27782780. [Google Scholar]
  9. Poon LL, Wong BW, Ma EH, Chan KH, Chow LM, Abeyewickreme W, Tangpukdee N, Yuen KW, Guan Y, Looareesuwan S, Peiris JS, , 2006. Sensitive and inexpensive molecular test for falciparum malaria: Detecting Plasmodium falciparum DNA directly from heat-treated blood by loop-mediated isothermal amplification. Clin Chem 52: 303306.[Crossref] [Google Scholar]
  10. Nagamine K, Hase T, Notomi T, , 2002. Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probes 16: 223229.[Crossref] [Google Scholar]
  11. Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T, , 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28: E63.[Crossref] [Google Scholar]
  12. Mori Y, Nagamine K, Tomita N, Notomi T, , 2001. Detection of loop-mediated isothermal reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun 289: 150154.[Crossref] [Google Scholar]
  13. Qiao Y-M, Guo Y-C, Zhang X-E, Zhou Y-F, Zhang Z-P, Wei H-P, Yang R-F, Wang D-B, , 2007. Loop-mediated isothermal amplification for rapid detection of Bacillus anthracis spores. Biotechnol Lett 29: 19391946.[Crossref] [Google Scholar]
  14. Tomita N, Mori Y, Kanda H, Notomi T, , 2008. Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products. Nat Protoc 3: 877882.[Crossref] [Google Scholar]
  15. Thekisoe OM, Kuboki N, Nambota A, Fujisaki K, Sugimoto C, Igarashi I, Yasuda J, Inoue N, , 2007. Species-specific loop-mediated isothermal amplification (LAMP) for diagnosis of trypanosomosis. Acta Trop 102: 182189.[Crossref] [Google Scholar]
  16. Sambrook J, Russell DW, Sambrook J, Russell DW, , 2001. Preparation and analysis of eukaryotic genomic DNA. , eds. Molecular Cloning. Third edition. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press, 6.16.30. [Google Scholar]
  17. Desquesnes M, Dávila AM, , 2002. Applications of PCR-based tools for detection and identification of animal trypanosomes: a review and perspectives. Vet Parasitol 109: 213231.[Crossref] [Google Scholar]
  18. Mori Y, Kitao M, Tomita N, Notomi T, , 2004. Real-time turbidimetry of LAMP reaction for quantifying template DNA. J Biochem Biophys Methods 59: 145157.[Crossref] [Google Scholar]
  19. Pineda V, Montalvo E, Alvarez D, Santamaria AM, Calzada JE, Saldana A, , 2008. Feeding sources and trypanosome infection index of Rhodnius pallescens in Chagas disease endemic area of Amador County, Panama. Rev Inst Med Trop Sao Paulo 50: 113116.[Crossref] [Google Scholar]
  20. Njiru ZK, Mikosza AS, Matovu E, Enyaru JC, Ouma JO, Kibona SN, Thompson RC, Ndung'u JM, , 2008. African trypanosomiasis: sensitive and rapid detection of subgenus Trypanozoon by loop-mediated isothermal amplification (LAMP) of parasite DNA. Int J Parasitol 38: 589599.[Crossref] [Google Scholar]
  21. Njiru ZK, Mikosza AS, Armstrong T, Enyaru JC, Ndung'u JM, Thompson AR, , 2008. Loop-mediated isothermal amplification (LAMP) method for rapid detection of Trypanosoma brucei rhodesiense . PLoS Negl Trop Dis 2: e147.[Crossref] [Google Scholar]
  22. Thekisoe OM, Omolo JD, Swai ES, Hayashida K, Zhang J, Sugimoto C, Inoue N, , 2007. Preliminary application and evaluation of loop-mediated isothermal amplification (LAMP) for detection of bovine theileriosis and trypanosomosis in Tanzania. Onderstepoort J Vet Res 74: 339342.[Crossref] [Google Scholar]
  23. Grab DJ, Lonsdale-Eccles J, Inoue N, , 2005. LAMP for tadpoles. Nat Methods 2: 635.[Crossref] [Google Scholar]
  24. Thekisoe OM, Bazie RS, Coronel-Servian AM, Sugimoto C, Kawazu S, Inoue N, , 2009. Stability of loop-mediated isothermal amplification (LAMP) reagents and its amplification efficiency on crude trypanosome DNA templates. J Vet Med Sci 71: 471475.[Crossref] [Google Scholar]

Data & Media loading...

  • Received : 09 Sep 2009
  • Accepted : 08 Feb 2010
  • Published online : 05 May 2010

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