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

• Tipo: Artigo em Revista Científica Internacional
• Data: 2012

Autores

Catarina Churro (Autor) Outra; Paulo Pereira (Autor) Outra; Vitor Vasconcelos (Autor) FCUP; Elisabete Valerio (Autor) Outra

Revista

Título: Archives of Microbiology

Vol. 194

Páginas: 749-757

ISSN: 0302-8933

Editora: Springer Nature

Indexação

ISI Web of Knowledge - 26 Citações

Scopus - 29 Citações

Classificação Científica

FOS: Ciências exactas e naturais > Ciências biológicas

Outras Informações

ID Authenticus: 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.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 9