TY - JOUR
T1 - Identification of Acanthamoeba at the Generic and Specific Levels Using the Polymerase Chain Reaction
AU - VODKIN, MICHAEL H.
AU - HOWE, DANIEL K.
AU - VISVESVARA, GOVINDA S.
AU - McLAUGHLIN, GERALD L.
N1 - Copyright:
Copyright 2016 Elsevier B.V., All rights reserved.
PY - 1992/5
Y1 - 1992/5
N2 - ABSTRACT. 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 (<47° C) led to more than one band. This multiple banding pattern could reproducibly classify Acanthamoeba at the strain level and was, in certain cases, diagnostic for known pathogenic strains. However, these assays need to be further refined to make them relevant for clinical purposes.
AB - ABSTRACT. 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 (<47° C) led to more than one band. This multiple banding pattern could reproducibly classify Acanthamoeba at the strain level and was, in certain cases, diagnostic for known pathogenic strains. However, these assays need to be further refined to make them relevant for clinical purposes.
KW - Amoeba
KW - Naegleria
KW - human pathogen
KW - repetitive DNA
KW - ribosomal DNA
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U2 - 10.1111/j.1550-7408.1992.tb01467.x
DO - 10.1111/j.1550-7408.1992.tb01467.x
M3 - Article
C2 - 1640385
AN - SCOPUS:0026857739
SN - 0022-3921
VL - 39
SP - 378
EP - 385
JO - Journal of Eukaryotic Microbiology
JF - Journal of Eukaryotic Microbiology
IS - 3
ER -