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Journal of Clinical Microbiology, January 2005, p. 41-48, Vol. 43, No. 1
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.1.41-48.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Multispacer Typing Technique for Sequence-Based Typing of Bartonella quintana

Unité des Rickettsies CNRS UMR 6020, IFR 48, Faculté de Médecine de Marseille, Université de la Méditerranée, Marseille, France,¹ Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala, Sweden²

Received 1 July 2004/ Returned for modification 30 August 2004/ Accepted 20 September 2004

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.

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