Loop-Mediated Isothermal Amplification Method for a Differential Identification of Human Taenia Tapeworms

  • Yasuhito Sako
  • Agathe Nkouawa
  • Tetsuya Yanagida
  • Akira Ito
Part of the Methods in Molecular Biology book series (MIMB, volume 1039)


Loop-mediated isothermal amplification (LAMP), which employs a Bst DNA polymerase with strand-displacement activity and four primers (two inner primers and two outer primers) recognizing six distinct regions on the target DNA, is a highly sensitive, specific, simple, and rapid nucleotide amplification method. Moreover, because the Bst DNA polymerase resists much DNA polymerase inhibitors present in biological specimens, the LAMP method is suitable for the detection of infectious agents from clinical material such as fecal samples. Here, we describe the LAMP method which can differentially detect and identify human Taenia tapeworms, Taenia solium, T. saginata, and T. asiatica, using DNA specimens prepared from parasite tissue and human fecal sample.

Key words

Loop-mediated isothermal amplification Taenia solium T. saginata T. asiatica Differential detection Proglottid Fecal sample 


  1. 1.
    Ito A, Nakao M, Wandra T (2003) Human taeniasis and cysticercosis in Asia. Lancet 362:1918–1920PubMedCrossRefGoogle Scholar
  2. 2.
    Garcia HH, Gonzalez AE, Evans CA, Gilman RH (2003) Taenia solium cysticercosis. Lancet 362:547–556PubMedCrossRefGoogle Scholar
  3. 3.
    Schantz PM, Cruz M, Sarti E, Pawlowski Z (1993) Potential eradicability of Taeniasis and cysticercosis. Bull Pan Am Health Organ 27:397–403PubMedGoogle Scholar
  4. 4.
    Gonzalez LM, Montero E, Harrison LJ, Parkhouse RM, Garate T (2000) Differential diagnosis of Taenia saginata and Taenia solium infection by PCR. J Clin Microbiol 38:737–744PubMedGoogle Scholar
  5. 5.
    Yamasaki H, Allan JC, Sato MO, Nakao M, Sako Y, Nakaya K et al (2004) DNA differential diagnosis of taeniasis and cysticercosis by multiplex PCR. J Clin Microbiol 42:548–553PubMedCrossRefGoogle Scholar
  6. 6.
    Nunes CM, Dias AK, Dias FE, Aoki SM, de Paula HB, Lima LG et al (2005) Taenia saginata: differential diagnosis of human taeniasis by polymerase chain reaction-restriction fragment length polymorphism assay. Exp Parasitol 110:412–415PubMedCrossRefGoogle Scholar
  7. 7.
    Mayta H, Gilman RH, Prendergast E, Castillo JP, Tinoco YO, Garcia HH et al (2008) Nested PCR for specific diagnosis of Taenia solium taeniasis. J Clin Microbiol 46:286–289PubMedCrossRefGoogle Scholar
  8. 8.
    Abu AlSoud W, Radstrom P (2000) Effects of amplification facilitators on diagnostic PCR in the presence of blood, feces, and meat. J Clin Microbiol 38:4463–4470Google Scholar
  9. 9.
    Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N et al (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:E63PubMedCrossRefGoogle Scholar
  10. 10.
    Mori Y, Nagamine K, Tomita N, Notomi T (2001) Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun 289:150–154PubMedCrossRefGoogle Scholar
  11. 11.
    Bakheit MA, Torra D, Palomino LA, Thekisoe OM, Mbati PA, Ongerth J et al (2008) Sensitive and specific detection of Cryptosporidium species in PCR-negative samples by loop-mediated isothermal DNA amplification and confirmation of generated LAMP products by sequencing. Vet Parasitol 158:11–22PubMedCrossRefGoogle Scholar
  12. 12.
    Liang SY, Chan YH, Hsia KT, Lee JL, Kuo MC, Hwa KY et al (2009) Development of loop-mediated isothermal amplification assay for detection of Entamoeba histolytica. J Clin Microbiol 47:1892–1895PubMedCrossRefGoogle Scholar
  13. 13.
    Macuhova K, Kumagai T, Akao N, Ohta N (2010) Loop-mediated isothermal amplification assay for detection and discrimination of Toxocara canis and Toxocara cati eggs directly from sand samples. J Parasitol 96:1224–1227PubMedCrossRefGoogle Scholar
  14. 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 15:62–69PubMedCrossRefGoogle Scholar
  15. 15.
    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 47:168–174PubMedCrossRefGoogle Scholar
  16. 16.
    Nkouawa A, Sako Y, Li T, Chen X, Wandra T, Swastika IK et al (2010) Evaluation of a loop-mediated isothermal amplification method using fecal specimens for differential detection of Taenia species from humans. J Clin Microbiol 48:3350–3352PubMedCrossRefGoogle Scholar
  17. 17.
    Nunes CM, Lima LG, Manoel CS, Pereira RN, Nakano MM, Garcia JF (2006) Fecal specimens preparation methods for PCR diagnosis of human taeniosis. Rev Inst Med Trop Sao Paulo 48:45–47PubMedCrossRefGoogle Scholar
  18. 18.
    Nkouawa A, Sako Y, Li T, Chen X, Nakao M, Yanagida T et al (2012) A loop-mediated isothermal amplification method for a differential identification of Taenia tapeworms from human: application to a field survey. Parasitol Int 61:723–725PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Yasuhito Sako
    • 1
  • Agathe Nkouawa
    • 1
  • Tetsuya Yanagida
    • 1
  • Akira Ito
    • 1
  1. 1.Department of ParasitologyAsahikawa Medical UniversityAsahikawaJapan

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