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Open Access Highly Accessed Methodology article

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

Geoff Hide1*, Jacqueline M Hughes1 and Robert McNuff2

Author Affiliations

1 Centre for Parasite Biology, Molecular Epidemiology and Ecology, Biosciences Research Institute, School of Environment and Life Sciences, University of Salford, Salford, UK, M5 4WT

2 SCIENTO, 61 Bury Old Road, Whitefield, Manchester, UK, M45 6TB

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BMC Ecology 2003, 3:7  doi:10.1186/1472-6785-3-7

Published: 30 September 2003

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 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.