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A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper
Geoff Hide1, Jacqueline M Hughes, Robert McNuff
1Centre for Parasite Biology, Molecular Epidemiology and Ecology, Biosciences Research Institute, School of Environment and Life Sciences, University of Salford, UK, M5 4WT. g.hide@salford.ac.uk
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 alpha-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.
