BOOK
MT-PCR - A rapid, reliable and effective tool for assessing toxic algal blooms in Victorian water supplies
Aaron Jex | Louise Baker | Raechel Littman
(2015)
Additional Information
Book Details
Abstract
This project addressed a need for a sensitive, accurate and reliable testing method to aid assessment of the toxicity of algal blooms and assist water management. Increasingly, diagnostic dilemmas are resolved through the use of DNA-based technologies which often provide high sensitivity and specificity and are efficient both in terms of costs and time. However to date, no such test was available to the Victorian water industries.
This project sought to bridge this gap by developing an automated DNA-based diagnostic assay for cyanobacterial bloom assessment blooms in Victorian waters. The assay exceeds expectation in its ability to accurately quantify levels of toxigenic cyanobacteria in bloom samples, retains exceptionally high specificity and sensitivity and each assay out-performs common conventional PCR approaches established in the literature. Four toxigen assays (microcystin, nodularin, cylindrospermopsin and saxitoxin) were designed, tested and optimised.
This book is co-published with Water Research Australia
Authors: Aaron Jex, Louise Baker and Raechel Littman, University of Melbourne
Table of Contents
| Section Title | Page | Action | Price |
|---|---|---|---|
| Cover | Cover | ||
| CONTENTS | v | ||
| FIGURES | vi | ||
| TABLES | vi | ||
| ABBREVIATIONS | vi | ||
| 1 INTRODUCTION | 1 | ||
| 1.1 BACKGROUND AND RELEVANCE | 1 | ||
| 1.2 Objectives | 1 | ||
| 1.3 Specific Milestone Goals | 2 | ||
| 1.4 Literature Review | 2 | ||
| 1.4.1 Cyanobacterial and the importance of cyanobacterial blooms | 2 | ||
| 1.4.2 Molecular aspects of cyanobacterial toxin production | 3 | ||
| 1.4.3 Detection of toxic cyanobacterial blooms | 3 | ||
| 2 MATERIALS AND METHODS | 5 | ||
| 2.1 Sample collection, preparation and genomic extraction | 5 | ||
| 2.2 Target and primer selection and assay design | 5 | ||
| 2.3 Assessment of control and field samples by MT-PCR | 5 | ||
| 2.4 Determination of diagnostic sensitivity and specificity | 6 | ||
| 2.5 Assessing quantitative potential of each MT-PCR toxin-producing gene assay | 7 | ||
| 3 RESULTS AND DISCUSSION | 8 | ||
| 3.1 Results | 8 | ||
| 3.1.1 Assessment of MT-PCR specificity, sensitivity and quantitative potential using control material | 8 | ||
| 3.1.2 Evaluation of MT-PCR in field samples and determination of diagnostics specificity and sensitivity | 8 | ||
| 3.2 Discussion | 9 | ||
| 4 SUMMARY AND CONCLUSIONS | 11 | ||
| 4.1 General summary | 11 | ||
| 4.2 Concluding statement | 11 | ||
| 5 RECOMMENDATIONS | 12 | ||
| 6 ACKNOWLEDGEMENTS | 13 | ||
| 7 REFERENCES | 14 | ||
| 8 APPENDIX I | 20 |