1. PCR-based identification of Klebsiella pneumoniae subsp. rhinoscleromatis, the agent of rhinoscleroma
- Author
-
Cindy Fevre, Philippe J. Sansonetti, Régis Tournebize, Ana S. Almeida, Sylvain Brisse, Alexis Delétoile, Lionel Frangeul, Valérie Barbe, Virginie Passet, Génotypage des Pathogènes et Santé Publique (Plate-forme) (PF8), Institut Pasteur [Paris], Genoscope - Centre national de séquençage [Evry] (GENOSCOPE), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Intégration et Analyse Génomique (Plate-Forme 4) (PF4), Pathogénie Microbienne Moléculaire, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Chaire Microbiologie et Maladies infectieuses, Collège de France (CdF (institution)), This work was supported financially by Institut Pasteur and the Genoscope (Evry, France), as well as by a generous gift from the Conny-Maeva Charitable Foundation., Institut Pasteur [Paris] (IP), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Collège de France - Chaire Microbiologie et Maladies infectieuses, and Autard, Delphine
- Subjects
Serotype ,Bacterial capsule ,Klebsiella ,MESH: Sequence Analysis, DNA ,Klebsiella pneumoniae ,MESH: Klebsiella pneumoniae ,MESH: Molecular Diagnostic Techniques ,MESH: Bacteriological Techniques ,Polymerase Chain Reaction ,law.invention ,Infectious Diseases/Bacterial Infections ,law ,[SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases ,MESH: Child ,Gene cluster ,Child ,MESH: Bacterial Proteins ,Polymerase chain reaction ,0303 health sciences ,Microbiology/Microbial Evolution and Genomics ,biology ,lcsh:Public aspects of medicine ,MESH: Polymorphism, Single Nucleotide ,Rhinoscleroma ,3. Good health ,Infectious Diseases ,Molecular Diagnostic Techniques ,Multigene Family ,[SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases ,MESH: Rhinoscleroma ,Microbiology/Microbial Physiology and Metabolism ,Female ,Research Article ,DNA, Bacterial ,MESH: DNA Primers ,lcsh:Arctic medicine. Tropical medicine ,lcsh:RC955-962 ,Molecular Sequence Data ,Porins ,Polymorphism, Single Nucleotide ,Sensitivity and Specificity ,Microbiology ,03 medical and health sciences ,Complete sequence ,Bacterial Proteins ,MESH: Bacterial Capsules ,medicine ,Humans ,Bacterial Capsules ,DNA Primers ,030304 developmental biology ,Bacteriological Techniques ,MESH: Humans ,MESH: Molecular Sequence Data ,MESH: Porins ,030306 microbiology ,Infectious Diseases/Respiratory Infections ,Public Health, Environmental and Occupational Health ,Microbiology/Medical Microbiology ,lcsh:RA1-1270 ,MESH: Polymerase Chain Reaction ,Sequence Analysis, DNA ,biology.organism_classification ,medicine.disease ,MESH: DNA, Bacterial ,MESH: Sensitivity and Specificity ,Infectious Diseases/Neglected Tropical Diseases ,MESH: Multigene Family ,MESH: Female - Abstract
Rhinoscleroma is a chronic granulomatous infection of the upper airways caused by the bacterium Klebsiella pneumoniae subsp. rhinoscleromatis. The disease is endemic in tropical and subtropical areas, but its diagnosis remains difficult. As a consequence, and despite available antibiotherapy, some patients evolve advanced stages that can lead to disfiguration, severe respiratory impairment and death by anoxia. Because identification of the etiologic agent is crucial for the definitive diagnosis of the disease, the aim of this study was to develop two simple PCR assays. We took advantage of the fact that all Klebsiella pneumoniae subsp. rhinoscleromatis isolates are (i) of capsular serotype K3; and (ii) belong to a single clone with diagnostic single nucleotide polymorphisms (SNP). The complete sequence of the genomic region comprising the capsular polysaccharide synthesis (cps) gene cluster was determined. Putative functions of the 21 genes identified were consistent with the structure of the K3 antigen. The K3-specific sequence of gene Kr11509 (wzy) was exploited to set up a PCR test, which was positive for 40 K3 strains but negative when assayed on the 76 other Klebsiella capsular types. Further, to discriminate Klebsiella pneumoniae subsp. rhinoscleromatis from other K3 Klebsiella strains, a specific PCR assay was developed based on diagnostic SNPs in the phosphate porin gene phoE. This work provides rapid and simple molecular tools to confirm the diagnostic of rhinoscleroma, which should improve patient care as well as knowledge on the prevalence and epidemiology of rhinoscleroma., Author Summary In humans, the bacterium Klebsiella pneumoniae subsp. rhinoscleromatis (also called clone Rhinoscleromatis, as it evolved from a single Klebsiella pneumoniae ancestral strain) causes rhinoscleroma, a chronic infection of the nose and throat. Identification of the bacterium from biopsies or nasal secretions is essential for diagnosis, and currently relies on the biochemical characteristics of clone Rhinoscleromatis and on detection of its capsule of antigenic type K3. Our aim was to develop two identification tests based on amplification by polymerase chain reaction (PCR) of specific portions of the genome of clone Rhinoscleromatis. We established the sequence of the capsular polysaccharide synthesis cluster and identified one gene sequence that was unique to K3 strains. A PCR test that targets this gene was shown to be specific for K3 strains. We also exploited unique DNA signatures of clone Rhinoscleromatis to develop a second PCR test, which is specific for this clone, thus allowing distinction from other K. pneumoniae K3 strains. These novel and simple identification tests should help to diagnose rhinoscleroma and to understand the epidemiology of this disease.
- Published
- 2011