Multispacer typing technique for sequence-based typing of Bartonella quintana

• Bartonella quintana is a worldwide fastidious bacterium of the Alphaproteobacteria responsible for bacillary angiomatosis, trench fever, chronic lymphadenopathy, and culture-negative endocarditis. The recent genome sequencing of a B. quintana isolate allowed us to propose a genome-wide sequence-based typing method. To ensure sequence discrimination based on highly polymorphic areas, we amplified and sequenced 34 spacers in a large collection of B. quintana isolates. Six of these exhibited polymorphisms and allowed the characterization of 4 genotypes. However, the strain variants suggested by the noncoding sequences did not correlate with the results of pulsed-field gel electrophoresis (PFGE), which suggested a higher degree of variability. Modification of the PFGE profile of one isolate after nine subcultures confirmed that rearrangement frequencies are high in this species, making PFGE unreliable for epidemiological purposes. The low extent of sequence heterogeneity in the species suggests a recent emergence of this bacterium as a human pathogen. Direct typing of natural samples allowed the identification of a fifth genotype in the DNA extracted from a human body louse collected in Burundi. We have named the typing technique herein described multispacer typing.

Ämnesord

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Nyckelord

Animals

Bacteremia/microbiology

Bacterial Typing Techniques

Bartonella quintana/*classification/genetics/isolation & purification

DNA; Bacterial/analysis

DNA; Ribosomal Spacer/*analysis

Electrophoresis; Gel; Pulsed-Field

Genotype

Heart Valves/microbiology

Humans

Lice/microbiology

Molecular Sequence Data

Research Support; Non-U.S. Gov't

Sequence Analysis; DNA

Trench Fever/microbiology

Molecular biology

Molekylärbiologi

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)