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Capture probe conjugated to paramagnetic nanoparticles for purification of Alexandrium species (Dinophyceae) DNA from environmental samples

Galluzzi, L., Bertozzini, E., Del Campo, A., Penna, A., Bruce, Ian J., Magnani, M. (2006) Capture probe conjugated to paramagnetic nanoparticles for purification of Alexandrium species (Dinophyceae) DNA from environmental samples. Journal of Applied Microbiology, 101 (1). pp. 36-43. ISSN 1364-5072. (doi:10.1111/j.1365-2672.2006.02952.x) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided) (KAR id:71952)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided. (Contact us about this Publication)
Official URL
http://dx.doi.org/10.1111/j.1365-2672.2006.02952.x

Abstract

Aims: To develop a rapid, cost-effective and selective Alexandrium DNA extraction procedure from environmental samples in order to provide good-quality template for the downstream PCR-based detection assay. Methods and Results: In this study, we tested a DNA extraction method based on silica-coated, superparamagnetic nanoparticles conjugated to a DNA-capture sequence (probe) complementary to a specific region of 5·8S rDNA of the genus Alexandrium. Cultured Alexandrium catenella cells were used as the harmful algal bloom species for the DNA extraction. Then, a PCR assay was performed with primers specific for the genus Alexandrium to assess the specificity and sensitivity of the nucleic acid extraction method. This method was applied to both cultured and field samples, reaching in both cases a detection limit of one A. catenella cell. Conclusions: The results suggest that the use of probe-conjugated paramagnetic nanoparticles could be effective for the specific purification of microalgal DNA in cultured or environmental samples, ensuring sensitivity and specificity of the subsequent PCR assays. Significance and Impact of the Study: The DNA extraction method optimized in this study represents a progress towards the rapid and efficient direct detection of Alexandrium cells in seawater monitoring. In fact, this method requires no other equipment than a magnet and a hybridization oven and, in principle, can be adapted to different toxic microalgal species and can be automated, allowing the processing of a high number of samples.

Item Type: Article
DOI/Identification number: 10.1111/j.1365-2672.2006.02952.x
Uncontrolled keywords: algal DNA; nanoparticle; ribosome DNA; sea water; silicon dioxide; toxin, algal bloom; capture method; detection method; dinoflagellate; DNA; microalga; seawater, Alexandrium catenella; algal bloom; article; controlled study; cost effectiveness analysis; Dinoflagellate; DNA extraction; DNA probe; DNA purification; DNA sequence; environmental monitoring; nonhuman; polymerase chain reaction; real time polymerase chain reaction; sensitivity and specificity, Animals; Dinoflagellida; DNA Primers; DNA, Algal; Environmental Monitoring; Eutrophication; Molecular Probe Techniques; Nanostructures; Nucleic Acid Hybridization; Phytoplankton; Reverse Transcriptase Polymerase Chain Reaction; Seawater; Sensitivity and Specificity; Water Microbiology, Alexandrium; Alexandrium catenella; algae; Dinophyceae
Subjects: Q Science > QR Microbiology
Divisions: Faculties > Sciences > School of Physical Sciences
Faculties > Sciences > School of Physical Sciences > Functional Materials Group
Depositing User: I. Bruce
Date Deposited: 01 Feb 2019 12:46 UTC
Last Modified: 30 May 2019 08:49 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/71952 (The current URI for this page, for reference purposes)
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