Cargando…

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 detect...

Descripción completa

Detalles Bibliográficos
Autores principales: Hide, Geoff, Hughes, Jacqueline M, McNuff, Robert
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2003
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC239857/
https://www.ncbi.nlm.nih.gov/pubmed/14516472
http://dx.doi.org/10.1186/1472-6785-3-7
_version_ 1782120980260323328
author Hide, Geoff
Hughes, Jacqueline M
McNuff, Robert
author_facet Hide, Geoff
Hughes, Jacqueline M
McNuff, Robert
author_sort Hide, Geoff
collection PubMed
description 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.
format Text
id pubmed-239857
institution National Center for Biotechnology Information
language English
publishDate 2003
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-2398572003-11-04 A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper Hide, Geoff Hughes, Jacqueline M McNuff, Robert BMC Ecol Methodology Article 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. BioMed Central 2003-09-30 /pmc/articles/PMC239857/ /pubmed/14516472 http://dx.doi.org/10.1186/1472-6785-3-7 Text en Copyright © 2003 Hide et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
spellingShingle Methodology Article
Hide, Geoff
Hughes, Jacqueline M
McNuff, Robert
A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper
title A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper
title_full A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper
title_fullStr A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper
title_full_unstemmed A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper
title_short A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper
title_sort rapid and simple method of detection of blepharisma japonicum using pcr and immobilisation on fta paper
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC239857/
https://www.ncbi.nlm.nih.gov/pubmed/14516472
http://dx.doi.org/10.1186/1472-6785-3-7
work_keys_str_mv AT hidegeoff arapidandsimplemethodofdetectionofblepharismajaponicumusingpcrandimmobilisationonftapaper
AT hughesjacquelinem arapidandsimplemethodofdetectionofblepharismajaponicumusingpcrandimmobilisationonftapaper
AT mcnuffrobert arapidandsimplemethodofdetectionofblepharismajaponicumusingpcrandimmobilisationonftapaper
AT hidegeoff rapidandsimplemethodofdetectionofblepharismajaponicumusingpcrandimmobilisationonftapaper
AT hughesjacquelinem rapidandsimplemethodofdetectionofblepharismajaponicumusingpcrandimmobilisationonftapaper
AT mcnuffrobert rapidandsimplemethodofdetectionofblepharismajaponicumusingpcrandimmobilisationonftapaper