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Rapid identification of Giardia duodenalis by loop-mediated isothermal amplification (LAMP) from faecal and environmental samples and comparative findings by PCR and real-time PCR methods

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Abstract

The detection of two human-pathogenic Giardia duodenalis assemblages A and B in faecal and water samples by loop-mediated isothermal amplification (LAMP) has been evaluated. The LAMP reaction is reproducible, rapid and specific for the detection G. duodenalis and has lower costs compared to the other molecular assays. This is the first application of LAMP for Giardia detection.

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References

  • Alhassan A, Thekisoe O, Yokoyama M, Inoue N, Motloang N, Mbati MY, Yin PA, Katayama H, Anzai Y, Sugimoto T, Igarashi I (2007) Development of loop-mediated isothermal amplification (LAMP) method for diagnosis of equine piroplasmosis. Vet Parasitol 143:155–160

    Article  PubMed  Google Scholar 

  • Amar CFL, Dear PH, McLauchlin J (2003) Detection and genotyping by real-time PCR/RFLP analyses of Giardia duodenalis from human faeces. J Med Microbiol 52:681–683

    Article  CAS  PubMed  Google Scholar 

  • Appelbee AJ, Frederick LM, Heitman TL, Olson ME (2003) Prevalence and genotyping of Giardia duodenalis from beef calves in Alberta, Canada. Vet Parasitol 112:289–294

    Article  CAS  PubMed  Google Scholar 

  • Bakheit A, Torra D, Palomino LA, Thekisoe O, Mbati PA, Ongerth J, Karanis P (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 158:11–22

    Article  CAS  PubMed  Google Scholar 

  • Boothroyd JC, Wang A, Campbell DA, Wang CC (1987) An unusually compact ribosomal DNA repeat in the protozoan Giardia lamblia. Nucleic Acids Res 15:4065–4084

    Article  CAS  PubMed  Google Scholar 

  • Dixon BR, Parenteau M, Martineau C, Fournier J (1997) A comparison of conventional microscopy, immunofluorescence microscopy and flow cytometry in the detection of Giardia lamblia cysts in beaver fecal samples. J Immunol Methods 202:27–33

    Article  CAS  PubMed  Google Scholar 

  • Hunter PR, Thompson RCA (2005) The zoonotic transmission of Giardia and Cryptosporidium. Int J Parasitol 35:1181–1190

    Article  PubMed  Google Scholar 

  • Ikadai H, Tanaka H, Shibahara N, Matsuu A, Uechi M, Itoh N, Oshiro S, Kudo N, Igarashi I, Oyamada T (2004) Molecular evidence of infections with Babesia gibsoni parasites in Japan and evaluation of the diagnostic potential of a loop-mediated isothermal amplification method. J Clin Microbiol 42:2465–2469

    Article  CAS  PubMed  Google Scholar 

  • Jakubowski W, Boutros S, Faber W, Fayer R, Ghiorse W, LeChevallier M, Rose J, Schaub S, Singh A, Stewart M (1996) Environmental methods for Cryptosporidium. J Am Water Works Assoc 88:107–121

    CAS  Google Scholar 

  • Karanis P, Kourenti C, Smith H (2007a) Water-borne transmission of protozoan parasites: a review of worldwide outbreaks and lessons we learnt. J Wat Health 5:1–38

    Article  Google Scholar 

  • Karanis P, Thekisoe O, Kiouptsi K, Ongerth J, Igarashi I, Inoue N (2007b) Development and preliminary evaluation of a loop-mediated isothermal amplification procedure for sensitive detection of Cryptosporidium oocysts in fecal and water samples. Appl Environ Microbiol 73:5660–5662

    Article  CAS  PubMed  Google Scholar 

  • Kuboki N, Inoue N, Sakurai T, Di Cello F, Grab DJ, Suzuki H, Sugimoto C, Igarashi I (2003) Loop-mediated isothermal amplification for detection of African trypanosomes. J Clin Microbiol 4:5517–5524

    Article  Google Scholar 

  • Medema G, Teunis P, Blokker M, Deere D, Davison A, Charles P, Loret JF (2006) WHO guidelines for drinking water quality, environmental health criteria, Cryptosporidium, Draft 2

  • 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 289:150–154

    Article  CAS  PubMed  Google Scholar 

  • Notomi T, Hiroto O, Harumi M, Toshihiro Y, Watanabe K, Nobuyuki A, Hase T (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:E63

    Article  CAS  PubMed  Google Scholar 

  • Ongerth JE, Stibbs HE (1987) Identification of Cryptosporidium in river water. Appl Environ Microbiol 53:672–676

    CAS  PubMed  Google Scholar 

  • Plutzer J, Karanis P, Domokos K, Törökné A, Márialigeti K (2008) Detection and characterization of Giardia and Cryptosporidium in Hungarian raw, surface and sewage water samples by IFT, PCR and sequence analysis of the SSUrRNA and GDH genes. Int J Hyg Environ Health 211:524–533

    Article  CAS  PubMed  Google Scholar 

  • Read CM, Monis PT, Thompson RCA (2004) Discrimination of all genotypes of Giardia duodenalis at the glutamate dehydrogenase locus using PCR-RFLP. Infect Gen Evol 4:125–130

    Article  CAS  Google Scholar 

  • Rimhanen-Finne R, Enemark HL, Kolehmainen J, Toropainen P, Hänninen ML (2007) Evaluation of immunofluorescence microscopy and enzyme-linked immunosorbent assay in detection of Cryptosporidium and Giardia infections in asymptomatic dogs. Vet Parasitol 145:345–348

    Article  CAS  PubMed  Google Scholar 

  • Savioli L, Smith H, Thompson RCA (2006) Giardia and Cryptosporidium join the ‘Neglected Diseases Initiative’. Trends Parasitol 22:203–208

    Article  CAS  PubMed  Google Scholar 

  • Sil AK, Das P, Bhattacharya S, Ghosh S, Chattopadhyay DJ (1998) Cloning of ribosomal RNA genes from an Indian isolate of Giardia lamblia and the use of intergenic non-transcribing spacer regions in the differentiation of Giardia from other enteric pathogens. J Biosci 23:557–564

    Article  CAS  Google Scholar 

  • Sotiriadou I, Karanis P (2008) Evaluation of loop-mediated isothermal amplification for detection of Toxoplasma gondii in water samples and comparative findings by polymerase chain reaction and immunofluorescence test. Diagn Microbiol Infect Dis 62:357–365

    Article  CAS  PubMed  Google Scholar 

  • Thekisoe O, Inoue N, Kuboki N, Tuntasuvan D, Bunnoy W, Borisutsuwan S, Igarashi I, Sugimoto C (2005) Evaluation of loop-mediated isothermal amplification (LAMP), PCR and parasitological tests for detection of Trypanosoma evansi in experimentally infected pigs. Vet Parasitol 130:327–330

    Article  CAS  PubMed  Google Scholar 

  • Thompson RCA (2004) The zoonotic significance and molecular epidemiology of Giardia and giardiasis. Vet Parasitol 126:15–35

    Article  PubMed  Google Scholar 

  • Traub RJ, Monis PT, Robertson I, Irwin P, Mencke N, Thompson RCA (2004) Epidemiological and molecular evidence supports the zoonotic transmission of Giardia among humans and dogs living in the same community. Parasitol 128:253–262

    Article  CAS  Google Scholar 

  • USEPA Method 1623 (2001) Cryptosporidium and Giardia in water by filtration/IMS/FA. EPA 821-R-01025. Office of Water 4603, U.S. Environmental Protection Agency, Washington, D.C. 3-5, 8, 31-32, 38--40

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Acknowledgements

This study was supported by Grant-in-Aid for Young Scientists, Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science and from the twenty-first Century COE Programme (A-1), Ministry of Education, Sports, Science, and Technology of Japan. We would like to thank Dr Jerry Ongerth for his editorial review and suggestions for the manuscript.

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Plutzer, J., Karanis, P. Rapid identification of Giardia duodenalis by loop-mediated isothermal amplification (LAMP) from faecal and environmental samples and comparative findings by PCR and real-time PCR methods. Parasitol Res 104, 1527–1533 (2009). https://doi.org/10.1007/s00436-009-1391-3

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  • DOI: https://doi.org/10.1007/s00436-009-1391-3

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