| Description | The aim of this project is to determine the origin of pathogens within drinking water biofilms. The project identifies analysis techniques and considers a range of possible environmental sources. Report first published in September 2005. |
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| ISBN | (Web Only) |
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| Official Print Publication Date | September 2005 |
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| Website Publication Date | April 01, 2008 |
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Belucha Sirerol, Sandra Lai, Stephanie Chisholm, Bengü Said, Gordon Nichols, Susanne Surman-Lee, Robert Owen, John V Lee.
ISBN 978 0 7559 1655 9 (Web only publication)
This document is also available in pdf format (1.8Mb)
Contents
2. SUMMARY
3. INTRODUCTION
4. MATERIALS AND METHODS.
4.1 SCOTTISH WATER RECEPTION AND PROCESSING
4.2 CULTURE
4.2.1 BATCH CULTURE
4.2.2 CONTINUOUS CULTURE
4.3 CONFIRMATION OF H. PYLORI BY BIOCHEMICAL TESTS AND GRAM STAIN
4.4 MOLECULAR DETECTION
4.4.1 DNA EXTRACTION METHODS
4.4.2 CONVENTIONAL PCR
4.4.3 REAL-TIME PCR LIGHTCYCLER
4.4.4 REAL-TIME PCR TAQMAN®
4.4.5 FLUORESCENT IN SITU HYBRIDISATION
5 RESULTS
5.1 COMPARISON OF DNA EXTRACTION METHODS.
5.2 DEVELOPMENT OF REAL-TIME PCR ASSAY FOR DETECTION OF H. PYLORI DIRECTLY FROM ENVIRONMENTAL SAMPLES
5.2.1 REAL-TIME PCR ASSAY USING LIGHTCYCLER TECHNOLOGY
5.2.2 REAL-TIME PCR ASSAY USING TAQMAN TECHNOLOGY
5.3 DETECTION AND QUANTIFICATION OF H. PYLORI BY FLUORESCENT IN SITU HYBRIDISATION
5.3.1 DETECTION AND QUANTIFICATION OF H. pylori.
5.3.2 OPTIMISATION OF FLUORESCENT IN SITU HYBRIDISATION ASSAY FOR H. pylori.
5.4 DETECTION OF H. PYLORI IN SCOTTISH MAINS WATER SAMPLES.
5.4.1 DETECTION OF H. pylori DIRECTLY FROM WATER BY CULTURE.
5.4.2 DETECTION OF H. pylori DIRECTLY FROM WATER BY CONVENTIONAL PCR.
5.4.3 DETECTION OF H. pylori DIRECTLY FROM SCOTTISH WATER BY TAQMAN® REAL-TIME PCR ASSAY
5.5 CHEMOSTAT
5.5.1 CHEMOSTAT PARAMETERS
5.5.2 H. pylori SURVIVAL IN SCOTTISH WATER IN THE CHEMOSTAT
5.5.3 MICROFLORA BACKGROUND
5.5.4 H. pylori SURVIVAL IN SCOTTISH WATER AGAINST MICROBIAL BACKGROUND IN PRESENCE OF GLASS TILES ONLY
5.5.5 H. pylori SURVIVAL IN SCOTTISH WATER AGAINST MICROBIAL BACKGROUND IN PRESENCE OF IRON TILES WITH GLASS TILES AS CONTROLS
5.5.6 COMPARISON OF RESULTS FROM SURVIVAL EXPERIMENTS
5.5.7 DETECTION OF H. pylori IN BIOFILMS.
6. DISCUSSION AND CONCLUSIONS
6.1 MOLECULAR TECHNIQUES TO DETECT H. PYLORI IN DRINKING WATER SAMPLES AND BIOFILMS.
6.1.1 DEVELOPMENT OF QUANTITATIVE REAL-TIME PCR ASSAY SPECIFIC FOR H. pylori.
6.1.2 DETECTION OF H. pylori IN DRINKING WATER AND BIOFILMS BY FLUORESCENCE IN SITU HYBRIDISATION.
6.2 DETECTION OF H. PYLORI H.PYLORI IN SCOTTISH DRINKING WATER
6.3 H. PYLORI SURVIVAL IN SCOTTISH DRINKING WATER
6.4 DETECTION OF H. PYLORI IN BIOFILMS.
7. FUTURE WORK
7.1 DETECTION OF H. PYLORI IN WATER BY MOLECULAR TECHNIQUES.
7.2 CHARACTERISATION OF SCOTTISH INDIGENOUS MICROFLORA.
7.3 FUTURE STUDIES ON H. PYLORI SURVIVAL IN WATER.
8 ACKNOWLEDGEMENTS
9 REFERENCES
10.1 APPENDIX A: MEDIA COMPOSITION
10.2 APPENDIX B: SOLUTIONS COMPOSITION
10.3 APPENDIX C: STANDARD QUANTIFICATION-DETAILED CALCULATIONS
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