Affiliation:
1. Research Laboratory for Infectious Diseases, National Institute of Public Health and the Environment, Bilthoven, The Netherlands
Abstract
ABSTRACT
A sensitive and specific PCR hybridization assay was developed for the simultaneous detection and identification of
Ehrlichia
and
Borrelia burgdorferi
sensu lato. In separate assays the 16S rRNA gene of
Ehrlichia
species and the 23S-5S rRNA spacer region of
B. burgdorferi
sensu lato were amplified and labeled by PCR. These PCR products were used in a reverse line blot hybridization assay in which oligonucleotide probes are covalently linked to a membrane in parallel lines. Hybridization of the samples with the oligonucleotide probes on this membrane enabled the simultaneous detection and identification of
Ehrlichia
,
B. burgdorferi
, and
Bartonella
species in 40 different samples. The application of the assay to DNA extracts from 121
Ixodes ricinus
ticks collected from roe deer demonstrated that 45% of these ticks carried
Ehrlichia
DNA. More than half of these positive ticks carried species with 16S rRNA gene sequences closely related to those of
E. phagocytophila
and the human granulocytic ehrlichiosis agent. The majority of the other positive ticks were infected with a newly identified
Ehrlichia
-like species. In addition, 13% of the ticks were infected with one or more
B. burgdorferi
genospecies. In more than 70% of the ticks 16S rRNA gene sequences for
Bartonella
species or other species closely related to
Bartonella
were found. In five of the ticks both
Ehrlichia
and
B. burgdorferi
species were detected.
Publisher
American Society for Microbiology
Cited by
389 articles.
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