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1.
Emerg Infect Dis ; 20(10): 1678-82, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25280366

RESUMO

Borrelia miyamotoi, a relapsing fever-related spirochete transmitted by Ixodes ticks, has been recently shown to be a human pathogen. To characterize the prevalence of this organism in questing Ixodes ticks, we tested 2,754 ticks for a variety of tickborne pathogens by PCR and electrospray-ionization mass spectrometry. Ticks were collected from California, New York, Connecticut, Pennsylvania, and Indiana in the United States and from Germany and the Czech Republic in Europe from 2008 through 2012. In addition, an isolate from Japan was characterized. We found 3 distinct genotypes, 1 for North America, 1 for Europe, and 1 for Japan. We found B. miyamotoi infection in ticks in 16 of the 26 sites surveyed, with infection prevalence as high as 15.4%. These results show the widespread distribution of the pathogen, indicating an exposure risk to humans in areas where Ixodes ticks reside.


Assuntos
Borrelia/classificação , Borrelia/isolamento & purificação , Ixodes/microbiologia , Animais , Borrelia/genética , Europa (Continente) , Genótipo , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Prevalência , Espectrometria de Massas por Ionização por Electrospray , Estados Unidos
2.
Vector Borne Zoonotic Dis ; 14(8): 584-91, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25072989

RESUMO

Abstract Ticks harbor numerous pathogens of significance to human and animal health. A better understanding of the pathogens carried by ticks in a given geographic area can alert health care providers of specific health risks leading to better diagnosis and treatments. In this study, we tested 226 Ixodes ricinis ticks from Southern Germany using a broad-range PCR and electrospray ionization mass spectrometry assay (PCR/ESI-MS) designed to identify tick-borne bacterial and protozoan pathogens in a single test. We found 21.2% of the ticks tested carried Borrelia burgdorferi sensu lato consisting of diverse genospecies; a surprisingly high percentage of ticks were infected with Babesia microti (3.5%). Other organisms found included Borrelia miyamotoi, Rickettsia helvetica, Rickettsia monacensis, and Anaplasma phagocytophilum. Of further significance was our finding that more than 7% of ticks were infected with more than one pathogen or putative pathogen.


Assuntos
Babesia microti/crescimento & desenvolvimento , Anaplasma phagocytophilum/genética , Anaplasma phagocytophilum/isolamento & purificação , Animais , Vetores Artrópodes/classificação , Vetores Artrópodes/crescimento & desenvolvimento , Vetores Artrópodes/microbiologia , Babesia/genética , Babesia/crescimento & desenvolvimento , Babesia/isolamento & purificação , Babesia microti/genética , Babesia microti/isolamento & purificação , Borrelia/genética , Borrelia/isolamento & purificação , DNA Bacteriano/genética , Alemanha/epidemiologia , Humanos , Ixodes/microbiologia , Ixodes/parasitologia , Reação em Cadeia da Polimerase/métodos , Prevalência , Rickettsia/genética , Rickettsia/isolamento & purificação , Espectrometria de Massas por Ionização por Electrospray/métodos , Doenças Transmitidas por Carrapatos
3.
Diagn Microbiol Infect Dis ; 76(4): 413-8, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23747029

RESUMO

A universal PCR and sequencing test, SepsiTest™ (Molzym, Germany) was evaluated for its applicability during daily diagnostic routine in a privately operated laboratory. In total, 96 specimens originating from 66 patients under suspect of infectious endocarditis, infections of joints, encephalitis/meningitis, systemic infections and infections of unknown genesis were PCR analysed and compared to culture results. Samples comprised cultured and non-cultured blood, synovial fluid, synovial tissue, heart valves, pacemakers, spinal tissue, cerebrospinal fluid, and swabs. PCR and culture were concordant in 26 negative and 8 positive cases (51.5%). A group of 25 patients was culture-negative but PCR-positive (37.9%). In at least 14 of these, common and/or rare aetiologies were identified, while for 4 patients the results of 16S PCR could not be unequivocally linked with the underlying disease. Benefits and limitations of the molecular test are discussed with special emphasis on technical and economic issues. In conclusion, SepsiTest™ proved to be a valuable tool for the diagnosis of aetiologies, particularly in cases of culture-negative patients who are under strong suspicion for an infection.


Assuntos
Candida/isolamento & purificação , Endocardite/diagnóstico , Bactérias Gram-Negativas/isolamento & purificação , Bactérias Gram-Positivas/isolamento & purificação , RNA Ribossômico 16S/genética , Kit de Reagentes para Diagnóstico , Sepse/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Técnicas de Tipagem Bacteriana , Candida/classificação , Candida/genética , Meios de Cultura , Endocardite/sangue , Endocardite/líquido cefalorraquidiano , Endocardite/microbiologia , Feminino , Bactérias Gram-Negativas/classificação , Bactérias Gram-Negativas/genética , Bactérias Gram-Positivas/classificação , Bactérias Gram-Positivas/genética , Valvas Cardíacas/microbiologia , Humanos , Articulações/microbiologia , Masculino , Pessoa de Meia-Idade , Técnicas de Tipagem Micológica , RNA Ribossômico 16S/classificação , Sensibilidade e Especificidade , Sepse/sangue , Sepse/líquido cefalorraquidiano , Sepse/microbiologia , Análise de Sequência de DNA , Líquido Sinovial/microbiologia
4.
Case Rep Med ; 2013: 610632, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24171004

RESUMO

Marine microorganisms are uncommon etiologies of skin and skin structure infections, that is, wound infections. We report a case of severe wound infection, caused by the marine Photobacterium damselae (Vibrionaceae), in a 64-year-old male patient, returning from Australia. The isolate tested positive for pPHDD1, a plasmid conferring high-level virulence. Furthermore, the wound was coinfected with Vibrio harveyi, a halophile bacterium, which has never been reported from human infections before. Identification was achieved by use of Matrix-Assisted Laser Desorption-Ionization Time of Flight Mass Spectrometry (MALDI-TOF) and confirmed by 16S rDNA sequencing. Data retrieval from bibliography was complicated since P. damselae has been renamed often with a number of synonyms present in the literature: Photobacterium damsela, Vibrio damselae, Vibrio damsela, Pasteurella damselae, and Listonella damsela. With all synonyms used as query terms, a literature search provided less than 20 cases published worldwide. A majority of those cases presenting as severe wound infection are even fatal following progression into necrotizing fasciitis. Management with daily wound dressing and antibiotic therapy (ofloxacin empirically, followed by doxycycline after availability of microbiology) led in the reported case to a favorable outcome, which seems to be, however, the exception based on a review of the available literature.

5.
Mol Cell Probes ; 19(4): 275-7, 2005 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16002259

RESUMO

We describe a case of a Mycobacterium isolated from a patient with cervical lymphadenitis which was initially identified by hsp65 RFLP as Mycobacterium marinum. Sequence analysis of the hsp65 DNA fragment and the 16S rDNA signature sequence, however, led to the identification of Mycobacterium malmoense. A point mutation in one of the restriction sites had shifted the M. malmoense typical RFLP pattern to the M. marinum specific RFLP pattern. As a consequence, care should be taken when identifying mycobacteria with the use of one molecular technique, only.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Proteínas de Choque Térmico/genética , Mycobacterium marinum/genética , Mycobacterium marinum/isolamento & purificação , Mycobacterium/genética , Mycobacterium/isolamento & purificação , Mutação Puntual/genética , Proteínas de Choque Térmico/química , Humanos , Peso Molecular , Reprodutibilidade dos Testes , Projetos de Pesquisa , Análise de Sequência de DNA
6.
Mol Cell Probes ; 18(1): 59-65, 2004 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-15036371

RESUMO

Analysis of heat shock protein 65 (hsp65) gene restriction fragment length polymorphism (RFLP) is done frequently to identify non-tuberculous mycobacteria (NTM) on a genetic basis. Here we report the results of analysing the hsp65 patterns of some rarely isolated NTM for which patterns have not been published before (Mycobacterium bohemicum, Mycobacterium hassiacum, Mycobacterium heckeshornense, Mycobacterium monacense, and Mycobacterium triplex). Furthermore new hsp65-variants for Mycobacterium interjectum (type II), Mycobacterium mucogenicum (type V), Mycobacterium gordonae (type VIII) and Mycobacterium paraffinicum (perhaps synonymous to Mycobacterium scrofulaceum) are described. All species were characterised by hsp65-RFLP, sequencing a 441-bp fragment of the hsp65 gene and sequencing the hypervariable region of the 16S rDNA. Additional data for less frequently isolated mycobacteria are provided and the hitherto described data for the Mycobacterium gordonae complex are summarised. Although the hsp65-RFLP analysis turned out to be a useful method a number of restraints (lack of standardisation, slight variability in fragment length) limits its broader use. Reliable identification of NTM needs, however, more than one molecular method. Identification results obtained by applying different methods yielded conflicting results. Confusion may be caused by older data base entries which are not updated and not longer reflect the actual systematic and taxonomy of the genus Mycobacterium.


Assuntos
Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana/métodos , Chaperoninas/genética , Mycobacterium/isolamento & purificação , Polimorfismo de Fragmento de Restrição , Sequência de Bases , Chaperonina 60 , Métodos , Mycobacterium/classificação , Mycobacterium/genética , RNA Ribossômico 16S/genética , Especificidade da Espécie
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