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Journal of Clinical Microbiology, October 2000, p. 3555-3560, Vol. 38, No. 10
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Brachyspira aalborgi Infection Diagnosed by Culture
and 16S Ribosomal DNA Sequencing Using Human Colonic Biopsy
Specimens
Wolfgang
Kraaz,1
Bertil
Pettersson,2
Ulf
Thunberg,3
Lars
Engstrand,4 and
Claes
Fellström5,*
Department of Pathology,1
Department of Oncology,3 and
Department of Microbiology,4 University
Hospital, S-751 85 Uppsala, Department of Biotechnology, Royal
Institute of Technology, S-100 44 Stockholm,2
and Department of Large Animal Clinical Sciences, Faculty of
Veterinary Medicine, Swedish University of Agricultural Sciences, S-750
07 Uppsala,5 Sweden
Received 17 March 2000/Returned for modification 11 June
2000/Accepted 28 July 2000
In this study we report on the isolation and characterization of
the intestinal spirochete Brachyspira aalborgi using human mucosal biopsy specimens taken from the colon of a young adult male
with intestinal spirochetosis. A selective medium, containing 400 µg
of spectinomycin/ml and 5 µg of polymyxin/ml was used for the
isolation procedure. A high degree of similarity, in terms of
phenotypic properties and 16S ribosomal DNA sequence, was observed between the isolated strain, named W1, and the type strain, 513A, of
B. aalborgi. A similarity of 99.7% in the nucleotide
sequence was found between W1 and 513AT, based on the
almost-complete gene. A short segment of the 16S rRNA gene was
amplified by PCR using genetic material enriched from paraffin-embedded
biopsy specimens, which were taken from the patient on two occasions.
The products showed 16S rRNA gene sequences virtually identical to that
of strain 513AT in the actual region. Immunohistochemistry
was performed on the colonic biopsy specimens with a polyclonal
antibody raised against an intestinal spirochete isolated in a previous
case of human intestinal spirochetosis. The antibody reacted strongly
with the spirochete on the luminal epithelium. No immune reaction was
seen within or below the surface epithelium. Routine histology did not
reveal signs of colitis. Electron microscopy showed spirochetes attached end-on to the colonic mucosal surface. The isolate grew poorly
on a commonly used selective medium for intestinal spirochetes, which
may explain previous failures to isolate B. aalborgi.
*
Corresponding author. Mailing address: Department of
Large Animal Clinical Sciences, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Box 7018, S-750 07 Uppsala, Sweden. Phone: 46 18 671473. Fax: 46 18 672919. E-mail:
Claes.Fellstrom{at}kirmed.slu.se.
Journal of Clinical Microbiology, October 2000, p. 3555-3560, Vol. 38, No. 10
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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