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 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.


CONCLUSIONS
This system has potential as a sensitive convenient detection system for Blepharisma and could be applied to other protozoan organisms.
D000818:Animals; D016798:Ciliophora / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000737:chemistry; D016054:DNA, Protozoan / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014404:Tubulin / Q000235:genetics; D014867:Water / Q000469:parasitology*
D017931:DNA Primers; D016054:DNA, Protozoan; D014404:Tubulin; D014867:Water