HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Al-Soud, W. A. Articles by Wadström, T. Search for Related Content PubMed PubMed Citation Articles by Al-Soud, W. A. Articles by Wadström, T. Agricola Articles by Al-Soud, W. A. Articles by Wadström, T.

Assessment of PCR-DGGE for the identification of diverse Helicobacter species, and application to faecal samples from zoo animals to determine Helicobacter prevalence

¹Department of Medical Microbiology, Dermatology and Infection, Lund University, Sölvegatan 23, SE-223 62 Lund, Sweden ²Department of Clinical Microbiology 7806, National University Hospital, Righospitalet, Tagensvej 20, DK-2200, Copenhagen, Denmark ³Danish Veterinary Institute, Bülowsvej 27, DK-1790, Copenhagen, Denmark ⁴Laboratorium voor Microbiologie, University of Ghent, Belgium

Correspondence Waleed Abu Al-Soud abu.al-soud{at}mmb.lu.se

Received March 19, 2003
Accepted May 14, 2003

Helicobacter species are fastidious bacterial pathogens that are difficult to culture by standard methods. A PCR-denaturing gradient gel electrophoresis (PCR-DGGE) technique for detection and identification of different Helicobacter species was developed and evaluated. The method involves PCR detection of Helicobacter DNA by genus-specific primers that target 16S rDNA and subsequent differentiation of Helicobacter PCR products by use of DGGE. Strains are identified by comparing mobilities of unknown samples to those determined for reference strains; sequence analysis can also be performed on purified amplicons. Sixteen DGGE profiles were derived from 44 type and reference strains of 20 Helicobacter species, indicating the potential of this approach for resolving infection of a single host by multiple Helicobacter species. Some more highly related species were not differentiated whereas in highly heterogeneous species, sequence divergence was observed and more than one PCR-DGGE profile was obtained. Application of the PCR-DGGE method to DNA extracted from faeces of zoo animals revealed the presence of Helicobacter DNA in 13 of 16 samples; a correlation was seen between the mobility of PCR products in DGGE analysis and DNA sequencing. In combination, this indicated that zoo animals are colonized by a wide range of different Helicobacter species; seven animals appeared to be colonized by multiple Helicobacter species. By this approach, presumptive identifications were made of Helicobacter bilis and Helicobacter hepaticus in a Nile crocodile, Helicobacter cinaedi in a baboon and a red panda, and Helicobacter felis in a wolf and a Taiwan beauty snake. All of these PCR products (400 bp) showed 100 % sequence similarity to 16S rDNA sequences of the mentioned species. These results demonstrate the potential of PCR-DGGE-based analysis for identification of Helicobacter species in complex ecosystems, such as the gastrointestinal tract, and could contribute to a better understanding of the ecology of helicobacters and other pathogens with a complex aetiology.

This article has been cited by other articles:

				H. Moyaert, F. Pasmans, R. Ducatelle, F. Haesebrouck, and M. Baele Evaluation of 16S rRNA Gene-Based PCR Assays for Genus-Level Identification of Helicobacter Species J. Clin. Microbiol., May 1, 2008; 46(5): 1867 - 1869. [Abstract] [Full Text] [PDF]

				T. Matsumoto, M. Goto, H. Murakami, T. Tanaka, H. Nishiyama, E. Ono, C. Okada, E. Sawabe, M. Yagoshi, A. Yoneyama, et al. Multicenter Study To Evaluate Bloodstream Infection by Helicobacter cinaedi in Japan J. Clin. Microbiol., September 1, 2007; 45(9): 2853 - 2857. [Abstract] [Full Text] [PDF]

				A. Garcia, S. Xu, F. E. Dewhirst, P. R. Nambiar, and J. G. Fox Enterohepatic Helicobacter species isolated from the ileum, liver and colon of a baboon with pancreatic islet amyloidosis. J. Med. Microbiol., November 1, 2006; 55(Pt 11): 1591 - 1595. [Abstract] [Full Text] [PDF]

				H.-O. Nilsson, I.-S. Ouis, U. Stenram, A. Ljungh, A. P. Moran, T. Wadstrom, and W. A. Al-Soud High Prevalence of Helicobacter Species Detected in Laboratory Mouse Strains by Multiplex PCR-Denaturing Gradient Gel Electrophoresis and Pyrosequencing J. Clin. Microbiol., August 1, 2004; 42(8): 3781 - 3788. [Abstract] [Full Text] [PDF]

				D. Asrat, I. Nilsson, Y. Mengistu, E. Kassa, S. Ashenafi, K. Ayenew, T. Wadstrom, and W. Abu-Al-Soud Prevalence of Helicobacter pylori vacA and cagA Genotypes in Ethiopian Dyspeptic Patients J. Clin. Microbiol., June 1, 2004; 42(6): 2682 - 2684. [Abstract] [Full Text] [PDF]