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 Shen, Z.
Right arrow Articles by Fox, J. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Shen, Z.
Right arrow Articles by Fox, J. G.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, September 1998, p. 2447-2453, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Development of a PCR-Restriction Fragment Length Polymorphism Assay Using the Nucleotide Sequence of the Helicobacter hepaticus Urease Structural Genes ureAB

Zeli Shen,1 David B. Schauer,1,2 Harry L. T. Mobley,3 and James G. Fox1,2,*

Divisions of Comparative Medicine1 and Toxicology,2 Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, and Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, Maryland 212013

Received 19 February 1998/Returned for modification 5 May 1998/Accepted 10 June 1998

Infection with Helicobacter hepaticus causes chronic active hepatitis in certain strains of mice and is associated with hepatocellular carcinoma in A/JCr mice. Like the gastric helicobacters, H. pylori and H. mustelae, H. hepaticus possesses a high level of urease activity. However, the H. hepaticus urease structural gene sequences have not been previously determined, and the role of the urease enzyme in colonization and in pathogenesis is not known. PCR was used to amplify a portion of the urease structural genes from H. hepaticus genomic DNA. Amplified DNA fragments were cloned, and the nucleotide sequence was determined. The deduced amino acid sequence of the partial H. hepaticus ureA gene product was found to exhibit 60% identity and 75% similarity to the predicted H. pylori UreA. The deduced amino acid sequence of a partial H. hepaticus ureB gene product exhibited 75% identity and 87% similarity to the predicted H. pylori UreB. Diversity among H. hepaticus isolates was evaluated by means of a restriction fragment length polymorphism (RFLP) assay. The 1.6-kb fragments within the ureAB open reading frames, amplified from 11 independent isolates, were digested with the restriction endonuclease HhaI. Three distinct RFLP patterns were observed. Identical RFLP profiles were noted in sequential isolates of one strain of H. hepaticus during an 18 month in vivo colonization study, suggesting that the urease genes of H. hepaticus are stable. The urease genes among H. hepaticus strains were also well conserved, showing 98.8 to 99% nucleotide sequence identity among three isolates analyzed. These findings indicate that H. hepaticus has urease structural genes which are homologous to those of the gastric Helicobacter species and that these gene sequences can be used in a PCR and RFLP assay for diagnosis of this important murine pathogen.

* Corresponding author. Mailing address: Division of Comparative Medicine, Massachusetts Institute of Technology, Bldg. 16 825C, 77 Massachusetts Ave., Cambridge, MA 02139. Phone: (617) 253-1757. Fax: (617) 258-5708. E-mail: jgfox{at}mit.edu.

Journal of Clinical Microbiology, September 1998, p. 2447-2453, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Feng, S., Ku, K., Hodzic, E., Lorenzana, E., Freet, K., Barthold, S. W. (2005). Differential Detection of Five Mouse-Infecting Helicobacter Species by Multiplex PCR. CVI 12: 531-536 [Abstract] [Full Text]  
  • Nyan, D. C., Welch, A. R., Dubois, A., Coleman, W. G. Jr. (2004). Development of a Noninvasive Method for Detecting and Monitoring the Time Course of Helicobacter pylori Infection. Infect. Immun. 72: 5358-5364 [Abstract] [Full Text]  
  • Avenaud, P., Le Bail, B., Mayo, K., Marais, A., Fawaz, R., Bioulac-Sage, P., Megraud, F. (2003). Natural History of Helicobacter hepaticus Infection in Conventional A/J Mice, with Special Reference to Liver Involvement. Infect. Immun. 71: 3667-3672 [Abstract] [Full Text]  
  • Beckwith, C. S., McGee, D. J., Mobley, H. L. T., Riley, L. K. (2001). Cloning, Expression, and Catalytic Activity of Helicobacter hepaticus Urease. Infect. Immun. 69: 5914-5920 [Abstract] [Full Text]  
  • Solnick, J. V., Schauer, D. B. (2001). Emergence of Diverse Helicobacter Species in the Pathogenesis of Gastric and Enterohepatic Diseases. Clin. Microbiol. Rev. 14: 59-97 [Abstract] [Full Text]  
  • Tanahashi, T., Kita, M., Kodama, T., Sawai, N., Yamaoka, Y., Mitsufuji, S., Katoh, F., Imanishi, J. (2000). Comparison of PCR-Restriction Fragment Length Polymorphism Analysis and PCR-Direct Sequencing Methods for Differentiating Helicobacter pylori ureB Gene Variants. J. Clin. Microbiol. 38: 165-169 [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»