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Infection and Immunity, October 2000, p. 6012-6026, Vol. 68, No. 10
Glaxo Wellcome Medicines Research Centre,
Stevenage SG2 1NY,1 and Department of
Medicine, University of Cambridge School of Clinical Medicine,
Addenbrookes Hospital, Cambridge CB2 2QQ,2
United Kingdom
Received 13 December 1999/Returned for modification 15 March
2000/Accepted 2 May 2000
Onset of rheumatoid arthritis (RA) is widely believed to be
preceded by exposure to some environmental trigger such as bacterial infectious agents. The influence of bacteria on RA disease onset or
pathology has to date been controversial, due to inconsistencies between groups in the report of bacterial species isolated from RA
disease tissue. Using a modified technique of reverse transcriptase-PCR amplification, we have detected bacterial rRNA in the synovial tissue
of late-stage RA and non-RA arthritis controls. This may be suggestive
of the presence of live bacteria. Sequencing of cloned complementary
rDNA (crDNA) products revealed a number of bacterial sequences in joint
tissue from each patient, and from these analyses a comprehensive
profile of the organisms present was compiled. This revealed a number
of different organisms in each patient, some of which are common to
both RA and non-RA controls and are probably opportunistic colonizers
of previously diseased tissue and others which are unique species.
These latter organisms may be candidates for a specific role in disease
pathology and require further investigation to exclude them as
causative agents in the complex bacterial millieu. In addition, many of
the detected bacterial species have not been identified previously from
synovial tissue or fluid from arthritis patients. These may not be
easily cultivable, since they were not revealed in previous studies
using conventional in vitro bacterial culture methods. In situ
hybridization analyses have revealed the joint-associated bacterial
rRNA to be both intra- and extracellular. The role of viable bacteria or their nucleic acids as triggers in disease onset or pathology in
either RA or non-RA arthritis controls is unclear and requires further investigation.
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Reverse Transcriptase-PCR Analysis of Bacterial
rRNA for Detection and Characterization of Bacterial Species in
Arthritis Synovial Tissue
*
Corresponding author. Mailing address: Glaxo Wellcome
Medicines Research Centre, Department of Immunopathology, Gunnels Wood Rd. Stevenage, Hertfordshire SG1 1NY, United Kingdom. Phone: (44) 1438 768027. Fax: (44) 1438 764818. E-mail:
kek23980{at}GlaxoWellcome.co.uk.
Present address: The Chester Beatty Institute for Cancer Research,
London SW3 6JB, United Kingdom.
Infection and Immunity, October 2000, p. 6012-6026, Vol. 68, No. 10
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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