Isolation of Chlamydia pneumoniae Clonal Variants by a Focus-Forming Assay

Author:

Gieffers Jens12,Belland Robert J.3,Whitmire William1,Ouellette Scot4,Crane Deborah1,Maass Matthias2,Byrne Gerald I.4,Caldwell Harlan D.1

Affiliation:

1. Laboratory of Intracellular Parasites

2. Institute of Medical Microbiology and Hygiene, Medical University of Luebeck, Luebeck, Germany

3. Laboratory of Human Bacterial Pathogenesis, National Institutes of AllergyInfectious Disease, National Institutes of Health, Hamilton, Montana 59840

4. Department of Medical Microbiology and Immunology, University of Wisconsin School of Medicine, Madison, Wisconsin 53711

Abstract

ABSTRACT Chlamydia pneumoniae is an obligate intracellular prokaryotic human pathogen that causes community-acquired respiratory infection and has been associated with atherosclerosis and cardiovascular disease. Unexpected results from genomic sequencing indicate that significant intrastrain polymorphism exists for some C. pneumoniae isolates. These polymorphisms could reflect genotypes with differing disease-causing characteristics. A definitive means to test this hypothesis is to obtain genetically homogeneous clonal populations of the pathogen and test them in models of infection and disease. To date, methods for cloning C. pneumoniae have not been reported. In this study, we describe the isolation of clonal variants with genetic differences in the tyrP locus from a polymorphic respiratory isolate, using a novel focus-forming assay. These results now allow investigations on the biology and pathogenesis of C. pneumoniae clonal genovars that could lead to new insights into the pathogenesis of this important human pathogen.

Publisher

American Society for Microbiology

Subject

Infectious Diseases,Immunology,Microbiology,Parasitology

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