Identification of Acanthamoeba at the Generic and Specific Levels Using the Polymerase Chain Reaction

We have adapted the polymerase chain reaction to identify strains of Acanthamoeba. Using computer‐assisted analysis, primers were designed from an anonymous repetitive sequence and from published sequences of 18S and 5S ribosomal RNA genes of A. castellanii. Amplification of a short ribosomal DNA ta...

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Veröffentlicht in:	The Journal of Protozoology 1992-05, Vol.39 (3), p.378-385
Hauptverfasser:	VODKIN, MICHAEL H., HOWE, DANIEL K., VISVESVARA, GOVINDA S., McLAUGHLIN, GERALD L.
Format:	Artikel
Sprache:	eng
Schlagworte:	Acanthamoeba Acanthamoeba - genetics Acanthamoeba - isolation & purification Amino Acid Sequence Amoeba Animals Base Sequence Biological and medical sciences DNA, Protozoan - genetics DNA, Ribosomal - genetics DNA, Single-Stranded - genetics General aspects human pathogen Infectious diseases Medical sciences Molecular Sequence Data Naegleria Parasitic diseases Polymerase Chain Reaction Protozoal diseases repetitive DNA Repetitive Sequences, Nucleic Acid ribosomal DNA
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Zusammenfassung:	We have adapted the polymerase chain reaction to identify strains of Acanthamoeba. Using computer‐assisted analysis, primers were designed from an anonymous repetitive sequence and from published sequences of 18S and 5S ribosomal RNA genes of A. castellanii. Amplification of a short ribosomal DNA target (272 base pairs) at restrictive annealing conditions (>50° C) resulted in a single band that was unique for the genus and distinguished Acanthamoeba from Naegleria. This assay functioned with fresh and formalin‐fixed cells as starting material. Amplification of longer targets (400–700 base pairs) at less restrictive annealing conditions (
ISSN:	0022-3921 1550-7408 2375-0804
DOI:	10.1111/j.1550-7408.1992.tb01467.x