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Journal of Clinical Microbiology, October 2008, p. 3270-3275, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00680-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Intragenomic Variation in the Internal Transcribed Spacer 1 Region of Dientamoeba fragilis as a Molecular Epidemiological Marker{triangledown}

Aldert Bart,1* Harold M. van der Heijden,2 Sophie Greve,1 Dave Speijer,3 Wil J. Landman,2 and Tom van Gool1

Section Parasitology, Department of Medical Microbiology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands,1 Animal Health Service Ltd. (GD), Deventer, The Netherlands,2 Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands3

Received 10 April 2008/ Returned for modification 18 June 2008/ Accepted 10 July 1008

Dientamoeba fragilis is a parasite that has been recognized to be a causative agent of gastrointestinal symptoms. Because in most studies only some infected persons experience symptoms, it is possible that D. fragilis is a heterogeneous species with variants that display similar morphologies but different pathogenicities. The search for genetic variation in D. fragilis was based on the small-subunit rRNA gene, which was not found to be useful for molecular epidemiology. In this report, we describe the isolation and characterization of additional rRNA gene cluster sequences, the internal transcribed spacer 1 (ITS-1)-5.8S rRNA gene-ITS-2 region. For comparative purposes, we also isolated the ITS-1-5.8S rRNA gene-ITS-2 region of Histomonas meleagridis, a protozoan parasite of birds and a close relative of D. fragilis. This region was found to be highly variable, and 11 different alleles of the ITS-1 sequence could be identified. Variation in the ITS-1 region was found to be intragenomic, with up to four different alleles in a single isolate. So-called C profiles were produced from the ITS-1 repertoire of single isolates,. Analysis of the C profiles of isolates from nonrelated patients identified several clearly distinguishable strains of D. fragilis. Within families, it was shown that members can be infected with the same or different strains of D. fragilis. In conclusion, the ITS-1 region can serve as a molecular epidemiological tool for the subtyping of D. fragilis directly from feces. This may serve as a means of studying the transmission, geographical distribution, and relationships between strains and the pathogenicity of this parasite.

* Corresponding author. Mailing address: Section Parasitology, Department of Medical Microbiology, University of Amsterdam, Meibergdreef 9, Amsterdam 1105 AZ, The Netherlands. Phone: 31-20-5663189. Fax: 31-20-5669207. E-mail: a.bart{at}amc.uva.nl

{triangledown} Published ahead of print on 23 July 2008.

Journal of Clinical Microbiology, October 2008, p. 3270-3275, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00680-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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