1887

Abstract

is associated with various human diseases such as periodontal disease and colorectal cancer (CRC); thus, detection might serve as a novel diagnostic tool. Here, we describe the development of a sensitive and rapid molecular method for detecting two genes: the highly conserved and , which encode a critical host colonization factor. Loop-mediated isothermal amplification (LAMP) primer sets for the rapid detection of and were designed and optimized. The primers yielded consistent negative results for 20 non- bacterial strains, confirming the high specificity of the primers. LAMP reaction primer sensitivity was determined, and its detection rate in comparison to conventional PCR was assessed using 57 clinical stool samples. The LAMP detection limit for and was 22.5 and 0.225 pg µl, respectively, indicating that the sensitivity of this method was 10-fold higher than that of conventional PCR. These results suggest that the LAMP technique is able to effectively identify via as well as detect its virulence factor. To the best of our knowledge, this study is the first to report the application of LAMP for the detection of and in . The LAMP method constitutes a sensitive and specific visual assay for the rapid detection of the pathogen .

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000300
2016-08-01
2021-08-02
Loading full text...

Full text loading...

/deliver/fulltext/jmm/65/8/760.html?itemId=/content/journal/jmm/10.1099/jmm.0.000300&mimeType=html&fmt=ahah

References

  1. Bauer C., Schoonbroodt D., Wagner C., Horsmans Y. 2000; Liver abscesses due to Fusobacterium species. Liver 20:267–268[PubMed] [CrossRef]
    [Google Scholar]
  2. Brook I. 1990; Bacteremia due to anaerobic bacteria in newborns. J Perinatol 10:351–356[PubMed]
    [Google Scholar]
  3. Castellarin M., Warren R. L., Freeman J. D., Dreolini L., Krzywinski M., Strauss J., Barnes R., Watson P., Allen-Vercoe E. et al. 2012; Fusobacterium nucleatum infection is prevalent in human colorectal carcinoma. Genome Res 22:299–306 [View Article][PubMed]
    [Google Scholar]
  4. Chen R., Tong Q., Zhang Y., Lou D., Kong Q., Lv S., Zhuo M., Wen L., Lu S. 2011; Loop-mediated isothermal amplification: rapid detection of Angiostrongylus cantonensis infection in Pomacea canaliculata . Parasites Vectors 4:204 [View Article]
    [Google Scholar]
  5. de Franchis R., Cross N. C., Foulkes N. S., Cox T. M. 1988; A potent inhibitor of Taq polymerase copurifies with human genomic DNA. Nucleic Acids Res 16:10355 [View Article][PubMed]
    [Google Scholar]
  6. Demling A., Demling C., Schwestka-Polly R., Stiesch M., Heuer W. 2009; Influence of lingual orthodontic therapy on microbial parameters and periodontal status in adults. Eur J Orthod 31:638–642 [View Article][PubMed]
    [Google Scholar]
  7. Flanagan L., Schmid J., Ebert M., Soucek P., Kunicka T., Liska V., Bruha J., Neary P., Dezeeuw N. et al. 2014; Fusobacterium nucleatum associates with stages of colorectal neoplasia development, colorectal cancer and disease outcome. Eur J Clin Microbiol Infect Dis 33:1381–1390 [View Article][PubMed]
    [Google Scholar]
  8. Han Y. W., Ikegami A., Rajanna C., Kawsar H. I., Zhou Y., Li M., Sojar H. T., Genco R. J., Kuramitsu H. K., Deng C. X. 2005; Identification and characterization of a novel adhesin unique to oral fusobacteria. J Bacteriol 187:5330–5340 [View Article][PubMed]
    [Google Scholar]
  9. Hara-Kudo Y., Yoshino M., Kojima T., Ikedo M. 2005; Loop-mediated isothermal amplification for the rapid detection of Salmonella . FEMS Microbiol Lett 253:155–161 [View Article][PubMed]
    [Google Scholar]
  10. Hirayama H., Kageyama S., Takahashi Y., Moriyasu S., Sawai K., Onoe S., Watanabe K., Kojiya S., Notomi T., Minamihashi A. 2006; Rapid sexing of water buffalo (Bubalus bubalis) embryos using loop-mediated isothermal amplification. Theriogenology 66:1249–1256 [View Article][PubMed]
    [Google Scholar]
  11. Kong Q. M., Lu S. H., Tong Q. B., Lou D., Chen R., Zheng B., Kumagai T., Wen L. Y., Ohta N., Zhou X. N. 2012; Loop-mediated isothermal amplification (LAMP): early detection of Toxoplasma gondii infection in mice. Parasit Vectors 5:2 [View Article][PubMed]
    [Google Scholar]
  12. Kostic A. D., Gevers D., Pedamallu C. S., Michaud M., Duke F., Earl A. M., Ojesina A. I., Jung J., Bass A. J. et al. 2012; Genomic analysis identifies association of Fusobacterium with colorectal carcinoma. Genome Res 22:292–298 [View Article][PubMed]
    [Google Scholar]
  13. Kostic A. D., Chun E., Robertson L., Glickman J. N., Gallini C. A., Michaud M., Clancy T. E., Chung D. C., Lochhead P. et al. 2013; Fusobacterium nucleatum potentiates intestinal tumorigenesis and modulates the tumor-immune microenvironment. Cell Host Microbe 14:207–215 [View Article][PubMed]
    [Google Scholar]
  14. Lin M., Liu W., Wang P., Tan J., Zhou Y., Wu P., Zhang T., Yuan J., Chen Y. 2015; Rapid detection of ermB gene in Clostridium difficile by loop-mediated isothermal amplification. J Med Microbiol 64:854–861 [View Article][PubMed]
    [Google Scholar]
  15. Loomer P. M. 2004; Microbiological diagnostic testing in the treatment of periodontal diseases. Periodontology 2000 34:49–56 [View Article]
    [Google Scholar]
  16. Mager D. L. 2006; Bacteria and cancer: cause, coincidence or cure? A review. J Transl Med 4:14 [View Article][PubMed]
    [Google Scholar]
  17. Marchesi J. R., Dutilh B. E., Hall N., Peters W. H., Roelofs R., Boleij A., Tjalsma H. 2011; Towards the human colorectal cancer microbiome. PLoS One 6:e20447 [View Article][PubMed]
    [Google Scholar]
  18. Ohtsuka K., Yanagawa K., Takatori K., Hara-Kudo Y. 2005; Detection of Salmonella enterica in naturally contaminated liquid eggs by loop-mediated isothermal amplification, and characterization of Salmonella isolates. Appl Environ Microbiol 71:6730–6735 [View Article][PubMed]
    [Google Scholar]
  19. Okafuji T., Yoshida N., Fujino M., Motegi Y., Ihara T., Ota Y., Notomi T., Nakayama T. 2005; Rapid diagnostic method for detection of mumps virus genome by loop-mediated isothermal amplification. J Clin Microbiol 43:1625–1631 [View Article][PubMed]
    [Google Scholar]
  20. Rowland I. R. 2009; The role of the gastrointestinal microbiota in colorectal cancer. Curr Pharm Des 15:1524–1527 [View Article][PubMed]
    [Google Scholar]
  21. Rubinstein M. R., Wang X., Liu W., Hao Y., Cai G., Han Y. W. 2013; Fusobacterium nucleatum promotes colorectal carcinogenesis by modulating E-cadherin/beta-catenin signaling via its FadA adhesin. Cell Host Microbe 14:195–206 [View Article][PubMed]
    [Google Scholar]
  22. Sklavounos A., Legakis N. J., Ioannidou H., Patrikiou A. 1986; Anaerobic bacteria in dentoalveolar abscesses. Int J Oral Maxillofac Surg 15:288–291 [View Article][PubMed]
    [Google Scholar]
  23. Song T., Toma C., Nakasone N., Iwanaga M. 2005; Sensitive and rapid detection of Shigella and enteroinvasive Escherichia coli by a loop-mediated isothermal amplification method. FEMS Microbiol Lett 243:259–263 [View Article][PubMed]
    [Google Scholar]
  24. Whitmore S. E., Lamont R. J. 2014; Oral bacteria and cancer. PLoS Pathog 10:e1003933 [View Article][PubMed]
    [Google Scholar]
  25. World Health Organization 2011; Fact sheet no. 297. World Health Organization, Geneva, Switzerland. http://www.who.int/mediacentre/factsheets/fs297/en/
  26. Wu N., Yang X., Zhang R., Li J., Xiao X., Hu Y., Chen Y., Yang F., Lu N. et al. 2013; Dysbiosis signature of fecal microbiota in colorectal cancer patients. Microb Ecol 66:462–470 [View Article][PubMed]
    [Google Scholar]
  27. Xu M., Yamada M., Li M., Liu H., Chen S. G., Han Y. W. 2007; FadA from Fusobacterium nucleatum utilizes both secreted and nonsecreted forms for functional oligomerization for attachment and invasion of host cells. J Biol Chem 282:25000–25009 [View Article][PubMed]
    [Google Scholar]
  28. Yang Z., Liu W., Cui Q., Niu W., Li H., Zhao X., Wei X., Wang X., Huang S. et al. 2014; Prevalence and detection of Stenotrophomonas maltophilia carrying metallo-beta-lactamase blaL1 in Beijing, China. Front Microbiol 5:692 [View Article][PubMed]
    [Google Scholar]
  29. Zhao X., He X., Li H., Zhao J., Huang S., Liu W., Wei X., Ding Y., Wang Z. et al. 2015; Detection of Streptococcus pyogenes using rapid visual molecular assay. FEMS Microbiol Lett 362:fnv148 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.000300
Loading
/content/journal/jmm/10.1099/jmm.0.000300
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF

Most cited this month Most Cited RSS feed

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