Affiliation:
1. Abteilung für medizinische Mikrobiologie und Hygiene, Universität Ulm, Ulm
2. Lehrstuhl für Mikrobiologie, Technische Universität München, Freising, Germany
Abstract
ABSTRACT
Chlamydiae are important pathogens of humans and animals but diagnosis of chlamydial infections is still hampered by inadequate detection methods. Fluorescence in situ hybridization (FISH) using rRNA-targeted oligonucleotide probes is widely used for the investigation of uncultured bacteria in complex microbial communities and has recently also been shown to be a valuable tool for the rapid detection of various bacterial pathogens in clinical specimens. Here we report on the development and evaluation of a hierarchic probe set for the specific detection and differentiation of chlamydiae, particularly
C. pneumoniae
,
C. trachomatis
,
C. psittaci
, and the recently described chlamydia-like bacteria comprising the novel genera
Neochlamydia
and
Parachlamydia
. The specificity of the nine newly developed probes was successfully demonstrated by in situ hybridization of experimentally infected amoebae and HeLa 229 cells, including HeLa 229 cells coinfected with
C. pneumoniae
and
C. trachomatis
. FISH reliably stained chlamydial inclusions as early as 12 h postinfection. The sensitivity of FISH was further confirmed by combination with direct fluorescence antibody staining. In contrast to previously established detection methods for chlamydiae, FISH was not susceptible to false-positive results and allows the detection of all recognized chlamydiae in one single step.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
67 articles.
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