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Development of an Acanthamoeba ‐specific Reverse Dot‐Blot and the Discovery of a New Ribotype
Author(s) -
CAST REBECCA J.
Publication year - 2001
Publication title -
journal of eukaryotic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 1066-5234
DOI - 10.1111/j.1550-7408.2001.tb00199.x
Subject(s) - acanthamoeba , biology , polymerase chain reaction , dot blot , ribosomal rna , identification (biology) , microbiology and biotechnology , protozoa , oligonucleotide , computational biology , gene , genetics , ecology
.Acanthamoeba is a genus of free‐living amoebae, of which some speeies have been found to cause opportunistic infections in humans. The identification of these amoebae in natural and disease samples is based primarily upon morphological features. While these features are more than adequate for identification to the genus level, they are not useful for species‐level identification. This not only leads to difficulty in the diagnosis of infections, but it makes an accurate assessment of the natural distribution of acanthamoebae very difficult to achieve. To improve this situation, a detection method was developed that utilizes both selective polymerase chain reaction amplification and the reverse dot‐blot. Oligonucleotides were designed to be specific for the described ribosomal groups (or ribotypes) of Acanthamoeba , as well as one specific for the genus itself. When this method was used to analyze a series of Acanthamoeba cultures from Pakistan, a new ribotype was identified in addition to the detection of the ubiquitously distributed T4 type.