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A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper

BACKGROUND: The rapid expansion in the availability of genome and DNA sequence information has opened up new possibilities for the development of methods for detecting free-living protozoa in environmental samples. The protozoan Blepharisma japonicum was used to investigate a rapid and simple detect...

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Detalles Bibliográficos
Autores principales: Hide, Geoff, Hughes, Jacqueline M, McNuff, Robert
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2003
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC239857/
https://www.ncbi.nlm.nih.gov/pubmed/14516472
http://dx.doi.org/10.1186/1472-6785-3-7
Descripción
Sumario:BACKGROUND: The rapid expansion in the availability of genome and DNA sequence information has opened up new possibilities for the development of methods for detecting free-living protozoa in environmental samples. The protozoan Blepharisma japonicum was used to investigate a rapid and simple detection system based on polymerase chain reaction amplification (PCR) from organisms immobilised on FTA paper. RESULTS: Using primers designed from the α-tubulin genes of Blepharisma, specific and sensitive detection to the equivalent of a single Blepharisma cell could be achieved. Similar detection levels were found using water samples, containing Blepharisma, which were dried onto Whatman FTA paper. CONCLUSION: This system has potential as a sensitive convenient detection system for Blepharisma and could be applied to other protozoan organisms.