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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 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.

SUBMITTER: Hide G 

PROVIDER: S-EPMC239857 | biostudies-literature | 2003 Sep

REPOSITORIES: biostudies-literature

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

Hide Geoff G   Hughes Jacqueline M JM   McNuff Robert R  

BMC ecology 20030930


<h4>Background</h4>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.<h4>Results</h4>Using primers designed from the alpha-tubulin genes of Blepharisma, spe  ...[more]

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