1887

Abstract

A loop-mediated isothermal amplification (LAMP) assay was developed to detect DNA for the first time from human faecal specimens. Four primers specific for were designed corresponding to small subunit rRNA gene sequences and tested on 100 human faecal specimens. Thirty-nine of the faecal specimens (39 %) were confirmed positive for by LAMP compared with 33 % by PCR and 32 % by light microscopy. LAMP yielded 94 % sensitivity and 88 % specificity compared with microscopy (sensitivity 48 %, specificity 76 %). No significant differences in positive detection of were found among the three methods (>0.05). However, LAMP has shown a substantial agreement with PCR (κ = 0.78) and fair agreement was demonstrated between microscopy and PCR (κ = 0.25). In conclusion, the LAMP assay proved to be useful as a simplified, rapid, sensitive and specific alternative molecular screening tool in the diagnosis of in faecal specimens.

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2015-11-01
2024-11-02
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References

  1. Bicart-Sée A., Massip P., Linas M.-D., Datry A. 2000; Successful treatment with nitazoxanide of Enterocytozoon bieneusi microsporidiosis in a patient with AIDS. Antimicrobial Agents and Chemotherapy. 44:167–168
    [Google Scholar]
  2. Botterel F., Minozzi C., Vittecoq D., Bourée P. 2002; Pulmonary localization of Enterocytozoon bieneusi in an AIDS patient: case report and review. J Clin Microbiol 40:4800–4801 [View Article][PubMed]
    [Google Scholar]
  3. Chabchoub N., Abdelmalek R., Mellouli F., Kanoun F., Thellier M., Bouratbine A., Aoun K. 2009; Genetic identification of intestinal microsporidia species in immunocompromised patients in Tunisia. Am J Trop Med Hyg 80:24–27[PubMed]
    [Google Scholar]
  4. da Silva A. J., Schwartz D. A., Visvesvara G. S., de Moura H., Slemenda S. B., Pieniazek N. J., Sensitive P. C. R. 1996; Diagnosis of infections by Enterocytozoon bieneusi (Microsporidia) using primers based on the region coding for small-subunit rRNA. J Clin Microbiol 34:986–987[PubMed]
    [Google Scholar]
  5. Didier E. S., Stovall M. E., Green L. C., Brindley P. J., Sestak K., Didier P. J. 2004; Epidemiology of microsporidiosis: sources and modes of transmission. Vet Parasitol 126:145–166 [View Article][PubMed]
    [Google Scholar]
  6. Endo S., Komori T., Ricci G., Sano A., Yokoyama K., Ohori A., Kamei K., Franco M., Miyaji M., Nishimura K. 2004; Detection of gp43 of Paracoccidioides brasiliensis by the loop-mediated isothermal amplification (LAMP) method. FEMS Microbiol Lett 234:93–97 [View Article][PubMed]
    [Google Scholar]
  7. Fakruddin M. 2011; Loop mediated isothermal amplification (LAMP) – an alternative to polymerase chain reaction (PCR). Bangladesh Res Publ J 5:1998–2003
    [Google Scholar]
  8. Franzen C., Müller A. 2001; Microsporidiosis: human diseases and diagnosis. Microbes Infect 3:389–400 [View Article][PubMed]
    [Google Scholar]
  9. Ghayour Najafabadi Z., Oormazdi H., Akhlaghi L., Meamar A. R., Nateghpour M., Farivar L., Razmjou E. 2014; Detection of Plasmodium vivax and Plasmodium falciparum DNA in human saliva and urine: loop-mediated isothermal amplification for malaria diagnosis. Acta Trop 136:44–49 [View Article][PubMed]
    [Google Scholar]
  10. Goodgame R., Stager C., Marcantel B., Alcocer E., Segura A. M. 1999; Intensity of infection in AIDS-related intestinal microsporidiosis. J Infect Dis 180:929–932 [View Article][PubMed]
    [Google Scholar]
  11. Khan M. G. M., Bhaskar K. R. H., Salam M. A., Akther T., Pluschke G., Mondal D. 2012; Diagnostic accuracy of loop-mediated isothermal amplification (LAMP) for detection of Leishmania DNA in buffy coat from visceral leishmaniasis patients. Parasit Vectors 5:280 [View Article][PubMed]
    [Google Scholar]
  12. Kumar S. S., Ananthan S., Joyee A. G. 2005; Detection of Enterocytozoon bieneusi (Microsporidia) by polymerase chain reaction (PCR) using species-specific primer in stool samples of HIV patients. Indian J Med Res 121:215–219[PubMed]
    [Google Scholar]
  13. Lau Y. L., Meganathan P., Sonaimuthu P., Thiruvengadam G., Nissapatorn V., Chen Y. 2010; Specific, sensitive, and rapid diagnosis of active toxoplasmosis by a loop-mediated isothermal amplification method using blood samples from patients. J Clin Microbiol 48:3698–3702 [View Article][PubMed]
    [Google Scholar]
  14. Lau Y. L., Fong M. Y., Mahmud R., Chang P. Y., Palaeya V., Cheong F. W., Chin L. C., Anthony C. N., Al-Mekhlafi A. M., Chen Y. 2011; Specific, sensitive and rapid detection of human Plasmodium knowlesi infection by loop-mediated isothermal amplification (LAMP) in blood samples. Malar J 10:197 [View Article][PubMed]
    [Google Scholar]
  15. Matos O., Lobo M. L., Xiao L. 2012; Epidemiology of Enterocytozoon bieneusi infection in humans. J Parasitol Res 2012:981424 [View Article][PubMed]
    [Google Scholar]
  16. 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 289:150–154 [View Article][PubMed]
    [Google Scholar]
  17. Müller A., Stellermann K., Hartmann P., Schrappe M., Fätkenheuer G., Salzberger B., Diehl V., Franzen C. 1999; A powerful DNA extraction method and PCR for detection of microsporidia in clinical stool specimens. Clin Diagn Lab Immunol 6243–246[PubMed]
    [Google Scholar]
  18. Nagamine K., Hase T., Notomi T. 2002; Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probes 16:223–229 [View Article][PubMed]
    [Google Scholar]
  19. Nagdev K. J., Kashyap R. S., Parida M. M., Kapgate R. C., Purohit H. J., Taori G. M., Daginawala H. F. 2011; Loop-mediated isothermal amplification for rapid and reliable diagnosis of tuberculous meningitis. J Clin Microbiol 49:1861–1865 [View Article][PubMed]
    [Google Scholar]
  20. 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 [View Article][PubMed]
    [Google Scholar]
  21. Parida M., Sannarangaiah S., Dash P. K., Rao P. V. L., Morita K. 2008; Loop mediated isothermal amplification (LAMP): a new generation of innovative gene amplification technique; perspectives in clinical diagnosis of infectious diseases. Rev Med Virol 18:407–421 [View Article][PubMed]
    [Google Scholar]
  22. 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 29:1939–1946 [View Article][PubMed]
    [Google Scholar]
  23. Qin C., Jian L., Xue-En F., Wei X. 2009; Rapid detection of porcine reproductive and respiratory syndrome virus by reverse transcription loop-mediated isothermal amplification. Intervirology 52:86–91 [View Article][PubMed]
    [Google Scholar]
  24. Rayan H. Z., Ismail O. A., El-Hamshary E. M., Ibrahim G. H., Omar S. M. 2013; Contribution of multiplex real time PCR for detection and differentiation of Enterocytozoon bieneusi and Encephalitozoon intestinalis in fecal samples of immunocompromised patients. Parasitol United J 6:51–58
    [Google Scholar]
  25. Salleh F. M., Al-Mekhlafi A. M., Nordin A., Yasin M., Al-Mekhlafi H. M., Moktar N. 2011; Evaluation of Gram-chromotrope Kinyoun staining technique: its effectiveness in detecting microsporidial spores in fecal specimens. Diagn Microbiol Infect Dis 69:82–85 [View Article][PubMed]
    [Google Scholar]
  26. 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 (IFT). Diagn Microbiol Infect Dis 62:357–365 [View Article][PubMed]
    [Google Scholar]
  27. Thekisoe O. M. M., Bazie R. S. B., Coronel-Servian A. M., 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 71:471–475 [View Article][PubMed]
    [Google Scholar]
  28. Verweij J. J., Ten Hove R., Brienen E. A. T., van Lieshout L. 2007; Multiplex detection of Enterocytozoon bieneusi and Encephalitozoon spp. in fecal samples using real-time PCR. Diagn Microbiol Infect Dis 57:163–167 [View Article][PubMed]
    [Google Scholar]
  29. Zhou D., Guo J., Xu L., Gao S., Lin Q., Wu Q., Wu L. Y. 2014; Establishment and application of a loop-mediated isothermal amplification (LAMP) system for detection of cry1Ac transgenic sugarcane. Sci Rep 44912[PubMed]
    [Google Scholar]
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