Capture probe conjugated to paramagnetic nanoparticles for purification of Alexandrium species (Dinophyceae) DNA from environmental samples
Article first published online: 16 MAY 2006
Journal of Applied Microbiology
Volume 101, Issue 1, pages 36–43, July 2006
How to Cite
Galluzzi, L., Bertozzini, E., Del Campo, A., Penna, A., Bruce, I.J. and Magnani, M. (2006), Capture probe conjugated to paramagnetic nanoparticles for purification of Alexandrium species (Dinophyceae) DNA from environmental samples. Journal of Applied Microbiology, 101: 36–43. doi: 10.1111/j.1365-2672.2006.02952.x
- Issue published online: 16 MAY 2006
- Article first published online: 16 MAY 2006
- 2005/0603: received 30 May 2005, revised 18 November 2005 and accepted 8 December 2005
- capture probe;
- paramagnetic nanoparticles;
- 5·8S rDNA
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.