This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Stinear, T.
Right arrow Articles by Johnson, P. D. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Stinear, T.
Right arrow Articles by Johnson, P. D. R.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, April 1999, p. 1018-1023, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Identification and Characterization of IS2404 and IS2606: Two Distinct Repeated Sequences for Detection of Mycobacterium ulcerans by PCR

Timothy Stinear,1,* Bruce C. Ross,2 John K. Davies,1 Lui Marino,3 Roy M. Robins-Browne,3 Frances Oppedisano,3 Aina Sievers,4 and Paul D. R. Johnson1,3,5

Department of Microbiology, Monash University, Clayton,1 Research and Development, CSL Limited,2 Department of Microbiology and Infectious Diseases, Royal Children's Hospital,3 Victorian Infectious Diseases Reference Laboratory,4 and Department of Infectious Diseases and Clinical Epidemiology, Monash Medical Centre,5 Victoria, Australia

Received 9 September 1998/Returned for modification 22 November 1998/Accepted 21 December 1998

Molecular analysis of Mycobacterium ulcerans has revealed two new insertion sequences (ISs), IS2404 and IS2606. IS2404 was identified by complete sequencing of a previously described repetitive DNA segment from M. ulcerans. This element is 1,274 bp long, contains 12-bp inverted repeats and a single open reading frame (ORF) potentially encoding a protein of 327 amino acids (aa), and apparently generates 7-bp direct repeats upon transposition. Amino acid similarity was found between the putative transposase and those encoded by ISs in other bacterial sequences from Aeromonas salmonicida (AsIs1), Escherichia coli (H repeat element), Vibrio cholerae (VcIS1), and Porphyromonas gingivalis (PGIS2). The second IS, IS2606, was discovered by sequence analysis of a HaeIII fragment of M. ulcerans genomic DNA containing a repetitive sequence. This element is 1,404 bp long, with 12-bp inverted repeats and a single ORF potentially encoding a protein of 445 aa. Database searches revealed a high degree of amino acid identity (70%) with the putative transposase of IS1554 from M. tuberculosis. Significant amino acid identity (40%) was also observed with transposases from several other microorganisms, including Rhizobium meliloti (ISRm3), Burkholderia cepacia (IS1356), Corynebacterium diphtheriae, and Yersinia pestis. PCR screening of DNA from 45 other species of mycobacteria with primers for IS2404 confirm that this element is found only in M. ulcerans. However, by PCR, IS2606 was also found in Mycobacterium lentiflavum, another slow-growing member of the genus Mycobacterium that is apparently genetically distinct from M. ulcerans. Testing the sensitivity of PCR based on IS2404 and IS2606 primers demonstrated the ability to detect 0.1 and 1 M. ulcerans genome equivalents, respectively. The ability to detect small numbers of cells by using two gene targets will be particularly useful for analyzing environmental samples, where there may be low concentrations of M. ulcerans among large numbers of other environmental mycobacteria.

* Corresponding author. Mailing address: Department of Microbiology, Monash University, Wellington Rd., Clayton 3168, Australia. Phone: 61 3 9905 4809. Fax: 61 3 9905 4811. E-mail: tim.stinear{at}med.monash.edu.au.

Journal of Clinical Microbiology, April 1999, p. 1018-1023, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Kaser, M., Hauser, J., Pluschke, G. (2009). Single Nucleotide Polymorphisms on the Road to Strain Differentiation in Mycobacterium ulcerans. J. Clin. Microbiol. 47: 3647-3652 [Abstract] [Full Text]  
  • Kaser, M., Ruf, M.-T., Hauser, J., Marsollier, L., Pluschke, G. (2009). Optimized Method for Preparation of DNA from Pathogenic and Environmental Mycobacteria. Appl. Environ. Microbiol. 75: 414-418 [Abstract] [Full Text]  
  • Chauty, A., Ardant, M.-F., Adeye, A., Euverte, H., Guedenon, A., Johnson, C., Aubry, J., Nuermberger, E., Grosset, J. (2007). Promising Clinical Efficacy of Streptomycin-Rifampin Combination for Treatment of Buruli Ulcer (Mycobacterium ulcerans Disease). Antimicrob. Agents Chemother. 51: 4029-4035 [Abstract] [Full Text]  
  • Fyfe, J. A. M., Lavender, C. J., Johnson, P. D. R., Globan, M., Sievers, A., Azuolas, J., Stinear, T. P. (2007). Development and Application of Two Multiplex Real-Time PCR Assays for the Detection of Mycobacterium ulcerans in Clinical and Environmental Samples. Appl. Environ. Microbiol. 73: 4733-4740 [Abstract] [Full Text]  
  • Yip, M. J., Porter, J. L., Fyfe, J. A. M., Lavender, C. J., Portaels, F., Rhodes, M., Kator, H., Colorni, A., Jenkin, G. A., Stinear, T. (2007). Evolution of Mycobacterium ulcerans and Other Mycolactone-Producing Mycobacteria from a Common Mycobacterium marinum Progenitor. J. Bacteriol. 189: 2021-2029 [Abstract] [Full Text]  
  • Stinear, T. P., Seemann, T., Pidot, S., Frigui, W., Reysset, G., Garnier, T., Meurice, G., Simon, D., Bouchier, C., Ma, L., Tichit, M., Porter, J. L., Ryan, J., Johnson, P. D.R., Davies, J. K., Jenkin, G. A., Small, P. L.C., Jones, L. M., Tekaia, F., Laval, F., Daffe, M., Parkhill, J., Cole, S. T. (2007). Reductive evolution and niche adaptation inferred from the genome of Mycobacterium ulcerans, the causative agent of Buruli ulcer. Genome Res 17: 192-200 [Abstract] [Full Text]  
  • WALSH, D. S., DELA CRUZ, E. C., ABALOS, R. M., TAN, E. V., WALSH, G. P., PORTAELS, F., MEYERS, W. M. (2007). CLINICAL AND HISTOLOGIC FEATURES OF SKIN LESIONS IN A CYNOMOLGUS MONKEY EXPERIMENTALLY INFECTED WITH MYCOBACTERIUM ULCERANS (BURULI ULCER) BY INTRADERMAL INOCULATION. Am J Trop Med Hyg 76: 132-134 [Abstract] [Full Text]  
  • Ranger, B. S., Mahrous, E. A., Mosi, L., Adusumilli, S., Lee, R. E., Colorni, A., Rhodes, M., Small, P. L. C. (2006). Globally Distributed Mycobacterial Fish Pathogens Produce a Novel Plasmid-Encoded Toxic Macrolide, Mycolactone F. Infect. Immun. 74: 6037-6045 [Abstract] [Full Text]  
  • Richter, E., Rusch-Gerdes, S., Hillemann, D. (2006). Evaluation of the GenoType Mycobacterium Assay for Identification of Mycobacterial Species from Cultures.. J. Clin. Microbiol. 44: 1769-1775 [Abstract] [Full Text]  
  • Hilty, M., Yeboah-Manu, D., Boakye, D., Mensah-Quainoo, E., Rondini, S., Schelling, E., Ofori-Adjei, D., Portaels, F., Zinsstag, J., Pluschke, G. (2006). Genetic Diversity in Mycobacterium ulcerans Isolates from Ghana Revealed by a Newly Identified Locus Containing a Variable Number of Tandem Repeats. J. Bacteriol. 188: 1462-1465 [Abstract] [Full Text]  
  • Ablordey, A., Hilty, M., Stragier, P., Swings, J., Portaels, F. (2005). Comparative Nucleotide Sequence Analysis of Polymorphic Variable-Number Tandem-Repeat Loci in Mycobacterium ulcerans. J. Clin. Microbiol. 43: 5281-5284 [Abstract] [Full Text]  
  • Phillips, R., Horsfield, C., Kuijper, S., Lartey, A., Tetteh, I., Etuaful, S., Nyamekye, B., Awuah, P., Nyarko, K. M., Osei-Sarpong, F., Lucas, S., Kolk, A. H. J., Wansbrough-Jones, M. (2005). Sensitivity of PCR Targeting the IS2404 Insertion Sequence of Mycobacterium ulcerans in an Assay Using Punch Biopsy Specimens for Diagnosis of Buruli Ulcer. J. Clin. Microbiol. 43: 3650-3656 [Abstract] [Full Text]  
  • WALSH, D. S., MEYERS, W. M., PORTAELS, F., LANE, J. E., MONGKOLSIRICHAIKUL, D., HUSSEM, K., GOSI, P., MYINT, K. S. A. (2005). HIGH RATES OF APOPTOSIS IN HUMAN MYCOBACTERIUM ULCERANS CULTURE-POSITIVE BURULI ULCER SKIN LESIONS. Am J Trop Med Hyg 73: 410-415 [Abstract] [Full Text]  
  • Mve-Obiang, A., Lee, R. E., Umstot, E. S., Trott, K. A., Grammer, T. C., Parker, J. M., Ranger, B. S., Grainger, R., Mahrous, E. A., Small, P. L. C. (2005). A Newly Discovered Mycobacterial Pathogen Isolated from Laboratory Colonies of Xenopus Species with Lethal Infections Produces a Novel Form of Mycolactone, the Mycobacterium ulcerans Macrolide Toxin. Infect. Immun. 73: 3307-3312 [Abstract] [Full Text]  
  • Rhodes, M. W., Kator, H., McNabb, A., Deshayes, C., Reyrat, J.-M., Brown-Elliott, B. A., Wallace, R. Jr, Trott, K. A., Parker, J. M., Lifland, B., Osterhout, G., Kaattari, I., Reece, K., Vogelbein, W., Ottinger, C. A. (2005). Mycobacterium pseudoshottsii sp. nov., a slowly growing chromogenic species isolated from Chesapeake Bay striped bass (Morone saxatilis). Int. J. Syst. Evol. Microbiol. 55: 1139-1147 [Abstract] [Full Text]  
  • Stinear, T. P., Pryor, M. J., Porter, J. L., Cole, S. T. (2005). Functional analysis and annotation of the virulence plasmid pMUM001 from Mycobacterium ulcerans. Microbiology 151: 683-692 [Abstract] [Full Text]  
  • Siegmund, V., Adjei, O., Racz, P., Berberich, C., Klutse, E., van Vloten, F., Kruppa, T., Fleischer, B., Bretzel, G. (2005). Dry-Reagent-Based PCR as a Novel Tool for Laboratory Confirmation of Clinically Diagnosed Mycobacterium ulcerans-Associated Disease in Areas in the Tropics Where M. ulcerans Is Endemic. J. Clin. Microbiol. 43: 271-276 [Abstract] [Full Text]  
  • Ablordey, A., Kotlowski, R., Swings, J., Portaels, F. (2005). PCR Amplification with Primers Based on IS2404 and GC-Rich Repeated Sequence Reveals Polymorphism in Mycobacterium ulcerans. J. Clin. Microbiol. 43: 448-451 [Abstract] [Full Text]  
  • Stinear, T. P., Mve-Obiang, A., Small, P. L. C., Frigui, W., Pryor, M. J., Brosch, R., Jenkin, G. A., Johnson, P. D. R., Davies, J. K., Lee, R. E., Adusumilli, S., Garnier, T., Haydock, S. F., Leadlay, P. F., Cole, S. T. (2004). From the Cover: Giant plasmid-encoded polyketide synthases produce the macrolide toxin of Mycobacterium ulcerans. Proc. Natl. Acad. Sci. USA 101: 1345-1349 [Abstract] [Full Text]  
  • Marsollier, L., Stinear, T., Aubry, J., Saint Andre, J. P., Robert, R., Legras, P., Manceau, A.-L., Audrain, C., Bourdon, S., Kouakou, H., Carbonnelle, B. (2004). Aquatic Plants Stimulate the Growth of and Biofilm Formation by Mycobacterium ulcerans in Axenic Culture and Harbor These Bacteria in the Environment. Appl. Environ. Microbiol. 70: 1097-1103 [Abstract] [Full Text]  
  • Daniel, A. K., Lee, R. E., Portaels, F., Small, P. L. C. (2004). Analysis of Mycobacterium Species for the Presence of a Macrolide Toxin, Mycolactone. Infect. Immun. 72: 123-132 [Abstract] [Full Text]  
  • Jenkin, G. A., Stinear, T. P., Johnson, P. D. R., Davies, J. K. (2003). Subtractive Hybridization Reveals a Type I Polyketide Synthase Locus Specific to Mycobacterium ulcerans. J. Bacteriol. 185: 6870-6882 [Abstract] [Full Text]  
  • Rondini, S., Mensah-Quainoo, E., Troll, H., Bodmer, T., Pluschke, G. (2003). Development and Application of Real-Time PCR Assay for Quantification of Mycobacterium ulcerans DNA. J. Clin. Microbiol. 41: 4231-4237 [Abstract] [Full Text]  
  • Marsollier, L., Robert, R., Aubry, J., Saint Andre, J.-P., Kouakou, H., Legras, P., Manceau, A.-L., Mahaza, C., Carbonnelle, B. (2002). Aquatic Insects as a Vector for Mycobacterium ulcerans. Appl. Environ. Microbiol. 68: 4623-4628 [Abstract] [Full Text]  
  • Chemlal, K., Huys, G., Laval, F., Vincent, V., Savage, C., Gutierrez, C., Laneelle, M.-A., Swings, J., Meyers, W. M., Daffe, M., Portaels, F. (2002). Characterization of an Unusual Mycobacterium: a Possible Missing Link between Mycobacterium marinum and Mycobacterium ulcerans. J. Clin. Microbiol. 40: 2370-2380 [Abstract] [Full Text]  
  • Russo, T. A., Carlino, U. B., Johnson, J. R. (2001). Identification of a New Iron-Regulated Virulence Gene, ireA, in an Extraintestinal Pathogenic Isolate of Escherichia coli. Infect. Immun. 69: 6209-6216 [Abstract] [Full Text]  
  • Chemlal, K., Huys, G., Fonteyne, P.-A., Vincent, V., Lopez, A. G., Rigouts, L., Swings, J., Meyers, W. M., Portaels, F. (2001). Evaluation of PCR-Restriction Profile Analysis and IS2404 Restriction Fragment Length Polymorphism and Amplified Fragment Length Polymorphism Fingerprinting for Identification and Typing of Mycobacterium ulcerans and M. marinum. J. Clin. Microbiol. 39: 3272-3278 [Abstract] [Full Text]  
  • Heckert, R. A., Elankumaran, S., Milani, A., Baya, A. (2001). Detection of a New Mycobacterium Species in Wild Striped Bass in the Chesapeake Bay. J. Clin. Microbiol. 39: 710-715 [Abstract] [Full Text]  
  • Stinear, T. P., Jenkin, G. A., Johnson, P. D. R., Davies, J. K. (2000). Comparative Genetic Analysis of Mycobacterium ulcerans and Mycobacterium marinum Reveals Evidence of Recent Divergence. J. Bacteriol. 182: 6322-6330 [Abstract] [Full Text]  
  • Stinear, T., Davies, J. K., Jenkin, G. A., Hayman, J. A., Oppedisano, F., Johnson, P. D. R. (2000). Identification of Mycobacterium ulcerans in the Environment from Regions in Southeast Australia in Which It Is Endemic with Sequence Capture-PCR. Appl. Environ. Microbiol. 66: 3206-3213 [Abstract] [Full Text]  
  • Stinear, T., Davies, J. K., Jenkin, G. A., Portaels, F., Ross, B. C., OppEdIsano, F., Purcell, M., Hayman, J. A., Johnson, P. D. R. (2000). A Simple PCR Method for Rapid Genotype Analysis of Mycobacterium ulcerans. J. Clin. Microbiol. 38: 1482-1487 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»