| 2870 A Molecular Analysis of the Bacteria Present within Oral Carcinoma | ||
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S.J. HOOPER1, S. CREAN2, M.J. FARDY1, M.A.O. LEWIS1, and M.J. WILSON1, 1Cardiff University, United Kingdom, 2North Glamorgan NHS Trust, Merthyr Tydfil, United Kingdom Objectives: In response to the increasing interest in the possible relationships between bacteria and the different stages of cancer development, the bacterial microbiota present within human oral squamous cell carcinoma (OSCC) was analysed using several molecular-based techniques. Methods: Tumourous and non-tumourous mucosal tissue specimens (approximately 1-cm3) were harvested from 10 OSCC patients at the time of surgery. Any microbial contamination on the specimens' surfaces was eliminated by immersion in Betadine® and washing with PBS. Bacteria were visualised within OSCC sections by performing fluorescent in situ hybridisation (FISH) with the universal oligonucleotide probe, EUB338-FITC. DNA was extracted from each aseptically macerated tissue specimen and used as template for PCR using primers targeting the ubiquitous 16S rRNA gene. Products from nested universal PCRs were separated by denaturing gradient gel electrophoresis (DGGE) in order to visualise the diversity of taxa within the specimens. Additionally, PCR products from three sets of primers, targeting Spirochaetes, Bacteroidetes and the domain Bacteria, were singularised by TA cloning and bacterial species were identified by partial sequencing of the 16S rRNA gene fragments. Results: Bacteria were detected within the tissue specimens by all three methods. A total of 70 distinct taxa were detected by PCR-cloning: 52 different phylotypes isolated from the tumourous tissues, and 37 taxa from within the non-tumourous specimens. Potentially significant differences between the composition of the microbiotas within the two tissue types were apparent and could be seen, though not quantified, by DGGE. Taxa isolated from within the tumour tissue were mostly from saccharolytic and aciduric genera, including Fusobacterium, Prevotella, Ralstonia and Streptococcus. Conclusion: The differences in the microbiota from tumourous and non-tumourous mucosae may indicate selective growth of bacteria within carcinoma tissue. Whether the presence of these bacteria within the mucosa has any bearing on the carcinogenic process is a concept worthy of further investigation.
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| Seq #294 - Immunology and Microbiology 10:45 AM-12:00 PM, Saturday, March 24, 2007 Ernest N. Morial Convention Center Exhibit Hall I2-J | ||
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