Fluorimetric Bioprobe Based on DNA-dye Intercalation - Application in Enviromental and Clinical Monitoring
(2007)- Abstract
- Nucleic acids have many unique structural features that offer reliable means of recognizing and monitoring many compounds of medical or environmental importance. In recent years, there has been an enormous increase in the use of nucleic acids as a tool to determine the chemical compounds of environmental and clinical interest and the major research activities in these areas have been directed towards the design of sequence-selective biosensors based on hybridization.
In this thesis, a novel bioassay method for detection of various substances with affinity for oligonucleotides including toxic aromatic compounds, metal ions, antibiotics and anticancer agents is described. The method is based on fluorimetric detection... (More) - Nucleic acids have many unique structural features that offer reliable means of recognizing and monitoring many compounds of medical or environmental importance. In recent years, there has been an enormous increase in the use of nucleic acids as a tool to determine the chemical compounds of environmental and clinical interest and the major research activities in these areas have been directed towards the design of sequence-selective biosensors based on hybridization.
In this thesis, a novel bioassay method for detection of various substances with affinity for oligonucleotides including toxic aromatic compounds, metal ions, antibiotics and anticancer agents is described. The method is based on fluorimetric detection wherein the long wavelength fluorescent intercalating dyes TO-PRO-3® or TOTO-3® and dsDNA form a complex. The presence of competing analytes disrupts this DNA-dye complex resulting in decreased fluorescence intensity. The analytes are quantitated indirectly by measuring the changes in the fluorescence intensity. A broad range of analytes like toxic aromatic compounds, metal ions, anticancer agents and antibiotics have been examined and quantified using this method. For example nano-molar concentrations of antibiotics like sulfathiazole and chloramphenicol could be detected; wide concentrations of clinically important anticancer agents, daunomycin and doxorubicin, were quantitated with good linearity.
Several home made fluorescence detection systems have been developed and used. They were based on optical fibers and different detection units like PMT or CCD camera. The assay throughput has been improved by the novel design of the detection system based on CCD camera.
Different sensitivities and detection ranges have been achieved by choosing different combinations of nucleic acids and analytes. This rapid and simple method has the potential for high throughput detection of environmental and clinical analytes. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/547790
- author
- Liu, Yang LU
- supervisor
- opponent
-
- Professor Guilbault, George, Department of Chemistry, University College Cork, Cork, Ireland
- organization
- publishing date
- 2007
- type
- Thesis
- publication status
- published
- subject
- keywords
- Bioteknik, Biotechnology, Biokemisk teknik, Biochemical technology, Antibiotics, Toxic aromatic compounds, Doxorubicin, High throughput screening, Fluorimetric assay, DNA
- publisher
- Lund University
- defense location
- Lecture hall B Center for Chemistry and Chemical Engineering, Sölvegatan 39, Lund
- defense date
- 2007-01-18 10:30:00
- ISBN
- 978-91-7422-138-1
- language
- English
- LU publication?
- yes
- additional info
- Yang Liu and Bengt Danielsson. 2005. Fluorimetric Broad-Range Screening of Compounds with Affinity for Nucleic Acids Analytical Chemistry, vol 77 pp 2450-2454. American Chemical SocietyYang Liu. 2006. Rapid Fluorometric Screening of Antibiotics in Seafood Microchimica Acta, vol 153 pp 133-137. SpringerWienNewYorkYang Liu. . Rapid High Throughput Assay for Fluorimetric Detection of Doxorubicin -Application of DNA-dye Bioprobe Elsevier B.V. (submitted)Yang Liu, Srimathi Soundararajan and Bengt Danielsson. . Development of DNA-Dye Complex Based Fluorimetric Bioprobe for Metal Ions Elsevier B.V. (submitted)
- id
- da1b8eb0-908c-47e5-b123-d925bc5c02ad (old id 547790)
- date added to LUP
- 2016-04-04 11:04:59
- date last changed
- 2018-11-21 21:02:33
@phdthesis{da1b8eb0-908c-47e5-b123-d925bc5c02ad, abstract = {{Nucleic acids have many unique structural features that offer reliable means of recognizing and monitoring many compounds of medical or environmental importance. In recent years, there has been an enormous increase in the use of nucleic acids as a tool to determine the chemical compounds of environmental and clinical interest and the major research activities in these areas have been directed towards the design of sequence-selective biosensors based on hybridization.<br/><br> <br/><br> In this thesis, a novel bioassay method for detection of various substances with affinity for oligonucleotides including toxic aromatic compounds, metal ions, antibiotics and anticancer agents is described. The method is based on fluorimetric detection wherein the long wavelength fluorescent intercalating dyes TO-PRO-3® or TOTO-3® and dsDNA form a complex. The presence of competing analytes disrupts this DNA-dye complex resulting in decreased fluorescence intensity. The analytes are quantitated indirectly by measuring the changes in the fluorescence intensity. A broad range of analytes like toxic aromatic compounds, metal ions, anticancer agents and antibiotics have been examined and quantified using this method. For example nano-molar concentrations of antibiotics like sulfathiazole and chloramphenicol could be detected; wide concentrations of clinically important anticancer agents, daunomycin and doxorubicin, were quantitated with good linearity.<br/><br> <br/><br> Several home made fluorescence detection systems have been developed and used. They were based on optical fibers and different detection units like PMT or CCD camera. The assay throughput has been improved by the novel design of the detection system based on CCD camera.<br/><br> <br/><br> Different sensitivities and detection ranges have been achieved by choosing different combinations of nucleic acids and analytes. This rapid and simple method has the potential for high throughput detection of environmental and clinical analytes.}}, author = {{Liu, Yang}}, isbn = {{978-91-7422-138-1}}, keywords = {{Bioteknik; Biotechnology; Biokemisk teknik; Biochemical technology; Antibiotics; Toxic aromatic compounds; Doxorubicin; High throughput screening; Fluorimetric assay; DNA}}, language = {{eng}}, publisher = {{Lund University}}, school = {{Lund University}}, title = {{Fluorimetric Bioprobe Based on DNA-dye Intercalation - Application in Enviromental and Clinical Monitoring}}, year = {{2007}}, }