A rapid and simple method of detection ofBlepharisma japonicumusing PCR and immobilisation on FTA paper

✍ Scribed by Geoff Hide; Jacqueline M Hughes; Robert McNuff

• Publisher: BioMed Central
• Year: 2003
• Tongue: English
• Weight: 700 KB
• Volume: 3
• Category: Article
• ISSN: 1472-6785
• DOI: 10.1186/1472-6785-3-7

✦ Synopsis

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.