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

Abstract

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 á-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. © 2003 Hide et al; licensee BioMed Central Ltd.