Species-specific real-time PCR cell number quantification of the bloom-forming cyanobacterium Planktothrix agardhii

• Type: Article in International Scientific Journal
• Date: 2012

Authors

Catarina Churro (Author) Other; Paulo Pereira (Author) Other; Vitor Vasconcelos (Author) FCUP; Elisabete Valerio (Author) Other

Journal

Title: Archives of Microbiology

Vol. 194

Pages: 749-757

ISSN: 0302-8933

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 26 Citations

Scopus - 29 Citations

Scientific classification

FOS: Natural sciences > Biological sciences

Other information

Authenticus ID: P-002-6JB

DOI: 10.1007/s00203-012-0809-y

Abstract (EN): A species-specific method to detect and quantify Planktothrix agardhii was developed by combining the SYBR Green I real-time polymerase chain reaction technique with a simplified DNA extraction procedure for standard curve preparation. Newly designed PCR primers were used to amplify a specific fragment within the rpoC1 gene. Since this gene exists in single copy in the genome, it allows the direct achievement of cell concentrations. The cell concentration determined by real-time PCR showed a linear correlation with the cell concentration determined from direct microscopic counts. The detection limit for cell quantification of the method was 8 cells mu L-1, corresponding to 32 cells per reaction. Furthermore, the real-time qPCR method described in this study allowed a successful quantification of P. agardhii from environmental water samples, showing that this protocol is an accurate and economic tool for a rapid absolute quantification of the potentially toxic cyanobacterium P. agardhii.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 9