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1.
J Appl Microbiol ; 121(3): 618-33, 2016 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-27321680

RESUMEN

Bacteriophages represent a simple viral model of basic research with many possibilities for practical application. Due to their ability to infect and kill bacteria, their potential in the treatment of bacterial infection has been examined since their discovery. With advances in molecular biology and gene engineering, the phage application spectrum has been expanded to various medical and biotechnological fields. The construction of bacteriophages with an extended host range or longer viability in the mammalian bloodstream enhances their potential as an alternative to conventional antibiotic treatment. Insertion of active depolymerase genes to their genomes can enforce the biofilm disposal. They can also be engineered to transfer various compounds to the eukaryotic organisms and the bacterial culture, applicable for the vaccine, drug or gene delivery. Phage recombinant lytic enzymes can be applied as enzybiotics in medicine as well as in biotechnology for pathogen detection or programmed cell death in bacterial expression strains. Besides, modified bacteriophages with high specificity can be applied as bioprobes in detection tools to estimate the presence of pathogens in food industry, or utilized in the control of food-borne pathogens as part of the constructed phage-based biosorbents.


Asunto(s)
Infecciones Bacterianas/tratamiento farmacológico , Bacteriófagos/genética , Terapia Biológica , Biotecnología/métodos , Microbiología Industrial , Animales , Bacterias/efectos de los fármacos , Biopelículas , Técnicas Biosensibles , Industria de Procesamiento de Alimentos , Ingeniería Genética , Humanos
2.
Folia Microbiol (Praha) ; 52(4): 331-8, 2007.
Artículo en Inglés | MEDLINE | ID: mdl-18062181

RESUMEN

Mutations extended the host range of the polyvalent bacteriophage 812 of the family Myoviridae in up to 95 % of Staphylococcus aureus strains and 43 % of strains of different coagulase-positive and -negative Staphylococcus species. Mutational changes in the genome of several host-range mutants of phage 812 were identified. Host-range mutant 812F1 harbors a deletion in endolysin gene that arose together with intron excision. Four mutants (812i, 812b, 812p, 812F3) harbor deletion in the structural gene orf8 that results from a genome rearrangement associated with intron insertion. This rearrangement was also detected in the genome of the closely related phages U16 and phi131. Another intron was discovered in the recA812 gene in these four mutants. An insertion was found in a non-coding region of the restriction fragment PstI-O of three mutants (812b, 812F3, 812g) and phages U16 and phi131. The above results contribute to the explanation of genetic factors affecting the host range of polyvalent staphylococcal bacteriophages.


Asunto(s)
Bacteriófagos/genética , Genoma Viral , Mutación , Staphylococcus aureus/virología , Secuencia de Aminoácidos , Secuencia de Bases , Endopeptidasas/química , Endopeptidasas/genética , Datos de Secuencia Molecular , Polimorfismo de Longitud del Fragmento de Restricción , ARN Viral/química , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Alineación de Secuencia , Proteínas de la Cola de los Virus/química , Proteínas de la Cola de los Virus/genética
3.
Klin Mikrobiol Infekc Lek ; 13(6): 231-5, 2007 Dec.
Artículo en Checo | MEDLINE | ID: mdl-18320502

RESUMEN

A solution to the problem of the increasing number of antibiotic-resistant bacterial strains can be the use of rational phage therapy. In the past, bacteriophages (phages) were often used for the treatment and prevention of infections and unlike antibiotic therapy, phage therapy caused almost no serious side effects. While previously several preparations containing whole phage particles were available for phage therapy, currently, the isolation of well characterised and purified phage components with antibacterial properties opens up new options for the management of intractable infections caused primarily by the bacterial genera Enterococcus, Escherichia, Klebsiella, Listeria, Proteus, Pseudomonas, Salmonella, Shigella, Staphylococcus and Streptococcus. In addition to human and veterinary medicine, the phage therapy principles also find use in the agriculture and food industry. Recent and former clinical studies as well as numerous animal model experiments have supported that phage therapy is an effective and safe alternative of antibiotic treatment of bacterial infections.


Asunto(s)
Infecciones Bacterianas/terapia , Bacteriófagos , Animales , Farmacorresistencia Microbiana , Humanos
4.
Sci Rep ; 7: 46319, 2017 04 13.
Artículo en Inglés | MEDLINE | ID: mdl-28406168

RESUMEN

Staphylococcus sciuri is a bacterial pathogen associated with infections in animals and humans, and represents a reservoir for the mecA gene encoding methicillin-resistance in staphylococci. No S. sciuri siphophages were known. Here the identification and characterization of two temperate S. sciuri phages from the Siphoviridae family designated ϕ575 and ϕ879 are presented. The phages have icosahedral heads and flexible noncontractile tails that end with a tail spike. The genomes of the phages are 42,160 and 41,448 bp long and encode 58 and 55 ORFs, respectively, arranged in functional modules. Their head-tail morphogenesis modules are similar to those of Staphylococcus aureus ϕ13-like serogroup F phages, suggesting their common evolutionary origin. The genome of phage ϕ575 harbours genes for staphylokinase and phospholipase that might enhance the virulence of the bacterial hosts. In addition both of the phages package a homologue of the mecA gene, which is a requirement for its lateral transfer. Phage ϕ879 transduces tetracycline and aminoglycoside pSTS7-like resistance plasmids from its host to other S. sciuri strains and to S. aureus. Furthermore, both of the phages efficiently adsorb to numerous staphylococcal species, indicating that they may contribute to interspecies horizontal gene transfer.


Asunto(s)
Genes Bacterianos , Metaloendopeptidasas/metabolismo , Fosfolipasas/metabolismo , Plásmidos/genética , Fagos de Staphylococcus/fisiología , Staphylococcus/virología , Transducción Genética , Transferencia de Gen Horizontal , Genoma Viral , Genómica/métodos , Especificidad del Huésped , Fagos de Staphylococcus/ultraestructura , Acoplamiento Viral
5.
Clin Microbiol Infect ; 12(4): 353-60, 2006 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-16524412

RESUMEN

This study describes the first molecular characterisation of clinical isolates of vancomycin-resistant enterococci (VRE) in the Czech Republic. Of 2647 patient isolates of Enterococcus spp. from 1997-2002, 121 (4.6%) were identified as VRE. The most common isolates were VanA+ Enterococcus faecium (78%) and VanB+ Enterococcus faecalis (10%). In addition, five VanA+ E. faecium isolates were obtained from environmental and staff sampling. Macrorestriction analysis of SmaI restriction fragment length polymorphism was performed for 54 VanA+ E. faecium clinical isolates and the five VanA+ E. faecium environmental isolates. Thirty-two unique restriction endonuclease patterns were identified, including two predominant clonal types represented by five or more isolates. Two environmental VanA+ E. faecium isolates were closely related to two patient isolates, which had an identical SmaI macrorestriction pattern. The results indicated potential survival of strains in the hospital environment and possible subsequent transmission to hospitalised patients.


Asunto(s)
Instituciones Oncológicas , Infección Hospitalaria/microbiología , Enterococcus faecium/genética , Enterococcus faecium/aislamiento & purificación , Infecciones por Bacterias Grampositivas/microbiología , Resistencia a la Vancomicina/genética , República Checa , Enterococcus faecalis/clasificación , Enterococcus faecalis/aislamiento & purificación , Enterococcus faecium/clasificación , Genotipo , Neoplasias Hematológicas , Humanos , Neoplasias , Polimorfismo de Longitud del Fragmento de Restricción
6.
Folia Microbiol (Praha) ; 50(6): 499-502, 2005.
Artículo en Inglés | MEDLINE | ID: mdl-16681147

RESUMEN

Rapid and specific detection of exfoliative toxin (ET)-producing Staphylococcus aureus strains by multiplex polymerase chain reaction (PCR) was used for identification of exfoliative toxin genes in a diverse set of 115 clinical S. aureus strains isolated in 14 Czech cities between 1998 and 2004. Fifty-nine wild-type ET-positive isolates of which 40 strains were the causative agents of toxic epidermolysis in neonates were classified into 4 PCR types. The genes coding for ETA, ETB or ETD were not detected in any of non-ET-producing isolates. The PCR method using the multiplex and specific primer set was shown to be reliable in rapid identification of the exfoliative toxin producing S. aureus and can be used as a convenient tool for hospital epidermolytic infection control.


Asunto(s)
Exfoliatinas/genética , Reacción en Cadena de la Polimerasa/métodos , Staphylococcus aureus/clasificación , Staphylococcus aureus/genética , Adulto , República Checa , ADN Bacteriano/análisis , Femenino , Humanos , Recién Nacido , Serotipificación , Infecciones Cutáneas Estafilocócicas/microbiología , Staphylococcus aureus/aislamiento & purificación
7.
FEMS Microbiol Lett ; 124(2): 131-9, 1994 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-7813882

RESUMEN

The genomes of 47 coagulase-negative staphylococcal strains assigned to different species were analysed by pulsed-field electrophoresis. The strains were clustered on the basis of their similarity in the SmaI restriction patterns into various groups, each group consisting of the type strain and the strains whose SmaI restriction patterns were similar to that of the type of strain. The SmaI restriction groups seem to correspond to the following species: Staphylococcus warneri, S. hominis, S. xylosus, S. lugdunensis, S. kloosii, S. haemolyticus, S. lentus, S. cohnii, S. equorum, S. chromogenes, S. saprophyticus, S. simulans, S. carnosus, S. capitis and S. auricularis. The species S. sciuri, S. caseolyticus, S. gallinarum, S. epidermidis and S. schleiferi were represented only by their type strains and showed no similarity in their SmaI restriction patterns neither to each other nor to all the other species investigated here. Thus, the classification of coagulase-negative staphylococcal strains into the above species seems to be confirmed also by genome restriction analysis carried out by pulsed-field gel electrophoresis.


Asunto(s)
ADN Bacteriano/análisis , Staphylococcus/genética , Enzimas de Restricción del ADN , Electroforesis en Gel de Campo Pulsado , Genoma , Mapeo Restrictivo
8.
FEMS Microbiol Lett ; 143(2-3): 203-10, 1996 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-8837473

RESUMEN

Several Staphylococcus aureus strains were lysogenized by the phages of serological group B (phages phi 53, phi 85) as well as by some of serological group F (phages phi 77, phi 84) and macrorestriction fragment patterns of genomic DNA were estimated in the lysogenized, non-lysogenic and delysogenized (cured of prophages) strains. It was shown that the integration of phage DNA into chromosome of S. aureus leads to specific changes in restriction fragment pattern in all the lysogenized strains. These changes correlate well with the SmaI restriction map of S. aureus NCTC 8325 since they concern the restriction fragments defined in this map. Phages phi 53 and phi 85 integrate into SmaI fragment B. On the other hand, phages phi 77 and phi 84 integrate into SmaI fragment E of the S. aureus restriction map. The prophages of strain NCTC 8511 have their integration sites, as follows: the phage designated by us phi M integrates in fragment A, whereas the integration site for phage phi J lies in fragment E. Phage phi M was estimated to be genetically related to phages of serological group A and phage phi J to those of serological group F. Evidence was given that lysogenization of S. aureus strains by at least four prophages does not cast any doubt upon the estimation of their genetic relatedness based on their similarity in restriction pattern.


Asunto(s)
Fagos de Staphylococcus/clasificación , Fagos de Staphylococcus/genética , Staphylococcus aureus/genética , Staphylococcus aureus/virología , Sitios de Ligazón Microbiológica/genética , ADN Bacteriano/genética , ADN Bacteriano/aislamiento & purificación , ADN Viral/genética , ADN Viral/aislamiento & purificación , Electroforesis en Gel de Campo Pulsado , Variación Genética , Lisogenia/genética , Mapeo Restrictivo , Serotipificación , Especificidad de la Especie
9.
FEMS Microbiol Lett ; 203(1): 23-7, 2001 Sep 11.
Artículo en Inglés | MEDLINE | ID: mdl-11557135

RESUMEN

Seven Enterococcus moraviensis and 16 Enterococcus haemoperoxidus as well as nine reference cultures of other enterococcal species obtained from the Czech Collection of Microorganisms were characterized using ribotyping with EcoRI and HindIII in the present work. The ribopatterns obtained by both restriction enzymes clearly distinguished all E. moraviensis and E. haemoperoxidus strains from the other enterococci (E. faecalis, E. faecium, E. avium, E. raffinosus, E. pseudoavium, E. malodoratus) and they differentiated both species from each other as well. Although all strains were isolated from different sampling sites, many strains shared the same band patterns. E. moraviensis formed four ribogroups using EcoRI and two ribogroups using HindIII restriction enzyme. E. haemoperoxidus gave six different patterns with EcoRI and five using the HindIII restriction enzyme.


Asunto(s)
Enterococcus/clasificación , Ribotipificación , ADN Bacteriano/análisis , Desoxirribonucleasa EcoRI/genética , Desoxirribonucleasa HindIII/genética , Enterococcus/genética , Homología de Secuencia de Ácido Nucleico , Especificidad de la Especie
10.
Folia Microbiol (Praha) ; 49(4): 353-86, 2004.
Artículo en Inglés | MEDLINE | ID: mdl-15530002

RESUMEN

Bacterial species of the genus Staphylococcus known as important human and animal pathogens are the cause of a number of severe infectious diseases. Apart from the major pathogen Staphylococcus aureus, other species until recently considered to be nonpathogenic may also be involved in serious infections. Rapid and accurate identification of the disease-causing agent is therefore prerequisite for disease control and epidemiological surveillance. Modern methods for identification and typing of bacterial species are based on genome analysis and have many advantages compared to phenotypic methods. The genotypic methods currently used in molecular diagnostics of staphylococcal species, particularly of S. aureus, are reviewed. Attention is also paid to new molecular methods with the highest discriminatory power. Efforts made to achieve interlaboratory reproducibility of diagnostic methods are presented.


Asunto(s)
Staphylococcus/genética , Técnicas de Tipificación Bacteriana , Genotipo , Humanos , Resistencia a la Meticilina , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN , Staphylococcus/clasificación , Staphylococcus/aislamiento & purificación
11.
Acta Virol ; 22(6): 443-50, 1978 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-35941

RESUMEN

Sedimentation of DNA of mycoplasma virus MV-Lg-L 172 in neutral and alkaline density gradients showed that the chromosome of this virus is most probably formed by a double-stranded circular DNA molecule with a molecular weight of approx. 10(7) daltons. These results were confirmed by direct electron microscopy of the viral DNA. Gamma-irradiation of the virus caused in its DNA single breaks on the average per each 66 eV of absorbed energy. Virus survival depended exponentially on the irradiation dose, D0 being between 60,000 and 70,000 rads. This dose induced on the average one single break in the DNA.


Asunto(s)
Bacteriófagos/análisis , ADN Circular/análisis , ADN Viral/análisis , Acholeplasma laidlawii , Bacteriófagos/efectos de la radiación , Radioisótopos de Cobalto , ADN Circular/efectos de la radiación , ADN Viral/efectos de la radiación , Microscopía Electrónica , Peso Molecular , Desnaturalización de Ácido Nucleico
12.
Int J Syst Bacteriol ; 49 Pt 3: 941-51, 1999 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-10425749

RESUMEN

On the basis of numerical analysis of 100 phenotypic features, the strains of two species, Staphylococcus carnosus and Staphylococcus piscifermentans, were differentiated into two separate phenons corresponding with the macrorestriction patterns of their genomic DNA, as well as with the results of ribotyping and PCR amplification of enterobacterial repetitive intergenic consensus sequences. One of the S. carnosus strains, the F-2 strain, was shown to be marginal, exhibiting the lowest genomic and phenotypic similarity to the S. carnosus type strain DSM 20501T. Two of the strains studied (strains S. carnosus SK 06 and S. piscifermentans SK 05) were phenotypically convergent, forming a separate phenon. They were phenotypically similar, even though the genomic DNA of one of them was homologous with that of the S. carnosus type strain, whereas that of the other was homologous with the genomic DNA of the S. piscifermentans type strain. In such cases, fingerprinting methods (particularly macrorestriction analysis and ribotyping) served as important correctives, as they allow phenotypically convergent strains to be distinguished on the basis of their genomic profiles. The results of this paper support the proposal for the new species Staphylococcus condimenti as well as the new subspecies Staphylococcus carnosus subsp. utilis.


Asunto(s)
Técnicas de Tipificación Bacteriana , Staphylococcus/clasificación , Staphylococcus/genética , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Bacteriano/aislamiento & purificación , ADN Ribosómico/genética , Desoxirribonucleasa EcoRI/metabolismo , Electroforesis en Gel de Campo Pulsado , Genotipo , Fenotipo , Reacción en Cadena de la Polimerasa/métodos , ARN Ribosómico/genética , Mapeo Restrictivo , Especificidad de la Especie
13.
Int J Syst Bacteriol ; 46(1): 216-22, 1996 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-8573498

RESUMEN

The genomic DNAs of 95 culture collection and hospital Staphylococcus aureus subsp. aureus strains of various origins, as well as the genomic DNAs of other coagulase-positive Staphylococcus species, were cleaved with restriction endonuclease SmaI and subjected to pulsed-field gel electrophoresis. The levels of similarity of the SmaI restriction patterns of the S. aureus subsp. aureus strains varied from 30 to 100%, which is considered characteristic of this species; thus, these organisms belonged to the same species restriction group. Within this range of similarity values 13 S. aureus intraspecies restriction groups were identified, and each group consisted of strains whose levels of similarity ranged from 65 to 100%. S. aureus subsp. aureus CCM 885T (T = type strain) belonged to the major intraspecies restriction group that comprised 39% of the S. aureus strains which we studied. The strains of the other coagulase-positive staphylococci, including Staphylococcus aureus subsp. anaerobius, Staphylococcus hyicus, Staphylococcus intermedius, Staphylococcus delphini, and Staphylococcus schleiferi subsp. coagulans, clustered with their type strains in separate restriction groups. S. aureus subsp. aureus exhibited almost no similarity to these species. We found 44-kb SmaI fragments in all of the S. aureus subsp. aureus and S. aureus subsp. anaerobius strains studied, and these fragments are considered characteristic of the species S. aureus. The high level of homology of these fragments was confirmed by the results of DNA hybridization experiments in which we used representatives of individual intraspecies restriction groups. Of the other staphylococci studied, only Staphylococcus epidermidis and one strain of S. hyicus contained these fragments. However, the levels of homology between these fragments and the fragments of S. aureus were found to be very low.


Asunto(s)
Variación Genética , Staphylococcus aureus/genética , Staphylococcus/genética , Coagulasa/metabolismo , ADN Bacteriano/genética , Desoxirribonucleasas de Localización Especificada Tipo II , Electroforesis en Gel de Campo Pulsado , Genoma Bacteriano , Filogenia , Mapeo Restrictivo , Staphylococcus/clasificación , Staphylococcus aureus/clasificación
14.
Arch Virol ; 149(9): 1689-703, 2004 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-15593413

RESUMEN

Conserved genomic sequences distinctive of Staphylococcus aureus phage types 3A, 11, 77, 187 and Twort, representative of phage serogroups A, B, F, L and D, were identified and characterized. PCR primers designed for the above sequences were used for development of a multiplex PCR assay which enabled us not only to classify all phages of the International Typing Set plus 16 additional phages, but also to detect prophages in S. aureus genomes. One to four different prophages were unambiguously detected in experimentally lysogenized S. aureus strains, and substantial variation in prophage content was found in 176 S. aureus clinical strains of different provenance. In addition, by using a comparative genomics approach, all the prophages in the S. aureus genomes sequenced to date could be revealed and classified.


Asunto(s)
Profagos/clasificación , Profagos/genética , Fagos de Staphylococcus/clasificación , Fagos de Staphylococcus/genética , Staphylococcus aureus/virología , Cartilla de ADN , ADN Viral/análisis , ADN Viral/química , ADN Viral/aislamiento & purificación , Genoma Viral , Lisogenia , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Profagos/aislamiento & purificación , Análisis de Secuencia de ADN , Fagos de Staphylococcus/aislamiento & purificación
15.
Electrophoresis ; 16(3): 366-76, 1995 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-7607171

RESUMEN

Seven representatives of the serogroup B Staphylococcus aureus bacteriophages, 29, 53, 55, 83A, 85, phi 11 and 80 alpha, were examined by capillary electrophoresis (CE) for genomic homology using DNA restriction analysis. Genomic DNA of individual bacteriophages was cleaved by HindIII restriction endonuclease, and the resulting restriction fragments were separated by standard horizontal agarose slab gel electrophoresis (SGE) as well as by CE in low-melting-point agarose solutions. The number and size of restriction fragments identified by both methods were compared. The high separation power of CE makes it possible to extend the restriction fragment patterns. In most of the restriction patterns, some additional restriction fragments as small as 150 bp, not identified by SGE, were detected. With respect to speed, high separation efficiency, low sample consumption and automation, CE offers a simple procedure for processing of multiple samples cost-effectively in a reasonable time. The comparison of the complemented restriction patterns of the different phage strains and the subsequent identification of their common fragments leads to a deeper understanding of their phylogenetic relationships. The genome homologies expressed for individual phage pairs in terms of coefficient F values ranged from 15 to 69%. These values are in good accordance with the degree of DNA homology of these phages as determined by DNA hybridization studies and thermal denaturation analysis of DNA by other authors. The total size of each phage genome was estimated by adding the sizes of individual restriction fragments.


Asunto(s)
Bacteriófagos/genética , Enzimas de Restricción del ADN , ADN Viral/análisis , Electroforesis en Gel de Agar/métodos , Staphylococcus aureus/virología , Acción Capilar , Desoxirribonucleasa EcoRI/metabolismo , Desoxirribonucleasa HindIII/metabolismo , Homología de Secuencia
16.
Electrophoresis ; 19(5): 695-700, 1998 May.
Artículo en Inglés | MEDLINE | ID: mdl-9629901

RESUMEN

The nucleotide sequence of a part of a 4.9 kbp common restriction fragment isolated from Staphylococcus aureus bacteriophage (bacterial virus) 3A has been determined by capillary electrophoresis (CE). The fast separation of sequencing fragments in linear polyacrylamide solution at a temperature of 55 degrees C allowed the reading of more than 650 bases of sequence in 60 min. The single strand (ss)DNA fragments were prepared by cycle sequencing with fluorescently labeled dideoxy-terminators on the cloned bacteriophage DNA template. With respect to analysis speed, sequence read-length, low sample consumption and automation, CE offers a simple, labor-saving and inexpensive procedure for DNA sequencing. Operating the CE columns at elevated temperature proved to be a rapid procedure capable of extending sequence read-length. The resulting sequence of the common restriction fragment can be used for the preparation of specific primers and oligonucleotide hybridization probes for identification of Staphylococcus aureus bacteriophages and/or prophages belonging to the bacteriophage species 3A.


Asunto(s)
ADN Viral/análisis , Electroforesis Capilar/métodos , Electroforesis en Gel de Poliacrilamida/métodos , Análisis de Secuencia de ADN/métodos , Fagos de Staphylococcus/genética , Resinas Acrílicas , Secuencia de Bases , ADN de Cadena Simple/análisis , Desoxirribonucleasa EcoRI , Colorantes Fluorescentes , Datos de Secuencia Molecular , Staphylococcus aureus/virología
17.
Mol Cell Probes ; 15(5): 249-57, 2001 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-11735296

RESUMEN

Primers were designed for polymerase chain reaction (PCR)-amplification of a genomic sequence specific to Staphylococcus aureus strains. The sequence corresponds to a part of the 44-kb Sma I fragment (fragment L on the S. aureus NCTC 8325 restriction map) which was found to be common to strains of the S. aureus species (Pantucek et al 1996, International Journal of Systematic Bacteriology, 46: 216-222). The labelled 44-kb Sma I restriction fragment derived from S. aureus NCTC 8325-4 was hybridized to the Eco RI restriction patterns of genomic DNA from 13 strains representing different macrorestriction types of S. aureus subsp. aureus. This made it possible to reveal the 2052 bp Eco RI restriction subfragment and to demonstrate its presence in all the tested strains. From the sequence of this subfragment, primers were designed by means of which the 826 bp amplicons were obtained in all 216 tested strains of S. aureus. No hybridization and PCR-products were observed in 40 collection strains of other staphylococcal species and subspecies as well as in 45 clinical strains of coagulase-negative staphylococci. These results lead us to the conclusion that the use of the above primers makes it possible to identify rapidly and reliably S. aureus strains of various provenance and different genotypes.


Asunto(s)
Desoxirribonucleasas de Localización Especificada Tipo II/metabolismo , Staphylococcus aureus/clasificación , Staphylococcus aureus/genética , Cartilla de ADN , Sondas de ADN , Genoma Bacteriano , Humanos , Hibridación de Ácido Nucleico , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Sensibilidad y Especificidad
18.
J Appl Microbiol ; 92(5): 951-7, 2002.
Artículo en Inglés | MEDLINE | ID: mdl-11972701

RESUMEN

AIMS: Enterococci associated with garden snails (Helix aspersa) were studied in order to obtain reliable species identification and characterization. METHODS AND RESULTS: Twelve yellow-pigmented and motile enterococci, isolated from the intestines of garden snails, were phenotypically close to Enterococcus casseliflavus, but they showed certain unusual biochemical characteristics. tRNA intergenic length polymorphism analysis (tDNA-PCR) divided all strains studied into two groups, in full agreement with biochemical test results. 16S rDNA sequencing, DNA base composition analysis and DNA-DNA hybridization results showed unambiguously that the enterococci studied belonged to the species Ent. casseliflavus. The representative strains of described ecovars were deposited in the Czech Collection of Microorganisms (CCM) as Ent. casseliflavus CCM 4868, 4869, 4870 and 4871. CONCLUSIONS: Enterococcus casseliflavus associated with garden snails can be subdivided into groups. SIGNIFICANCE AND IMPACT OF THE STUDY: Enterococcus casseliflavus differs from other enterococcal species in that it is typically associated with plants, soil, water and invertebrate animals. The different groups that can be found in these widely occurring bacteria are possibly source-specific ecovars, as exemplified by the Ent. casseliflavus inhabiting the intestines of snails.


Asunto(s)
Enterococcus/clasificación , Caracoles Helix/microbiología , Pigmentos Biológicos/metabolismo , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/análisis , ADN Bacteriano/química , ADN Bacteriano/genética , Enterococcus/genética , Enterococcus/aislamiento & purificación , Enterococcus/fisiología , Intestinos/microbiología , Movimiento , Hibridación de Ácido Nucleico , ARN Ribosómico 16S/genética , ARN de Transferencia/genética , Análisis de Secuencia de ADN
19.
Int J Syst Evol Microbiol ; 51(Pt 4): 1567-1574, 2001 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-11491359

RESUMEN

A polyphasic taxonomic approach was used to study atypical enterococci isolated from surface waters. All strains were characterized by physiological and biochemical tests as well as by genotyping. The results of biochemical tests and tRNA intergenic length polymorphism analysis (tDNA-PCR) divided all studied strains uniformly into two groups. Because these groups were clearly separated from all enterococcal species described to date, 16S rDNA sequence analysis, DNA base composition analysis and DNA-DNA hybridization of representative strains were done to elucidate the taxonomic position of the analysed groups. On the basis of the results obtained, the names Enterococcus haemoperoxidus (type strain CCM 4851T = LMG 19487T) and Enterococcus moraviensis (type strain CCM 4856T = LMG 19486T) are proposed for the two hitherto undescribed species. The type strains and reference cultures have been deposited in the Czech Collection of Microorganisms (CCM), Masaryk University, Brno, Czech Republic, and in the BCCM/LMG Culture Collection, Ghent University, Belgium.


Asunto(s)
Enterococcus/clasificación , Enterococcus/aislamiento & purificación , Microbiología del Agua , Composición de Base , Secuencia de Bases , República Checa , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/genética , Enterococcus/genética , Genotipo , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Fenotipo , Filogenia , Especificidad de la Especie , Terminología como Asunto
20.
Can J Microbiol ; 46(11): 1066-76, 2000 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-11109497

RESUMEN

On the basis of HindIII-restriction digest analysis of genomic DNAs, the S. aureus bacteriophages of the International Typing Set were divided into five clusters designated as A, F, Ba, Bb, and Bc. The clusters A and F include all the phages of serogroups A and F and correspond to species 3A and 77 proposed by Ackermann and DuBow (1987). On the other hand, the phages of serogroup B were divided into three clusters designated as Ba, Bb, and Bc that differ significantly each from the other in their restriction patterns. The clusters Ba and Bb may represent two separate species, while the cluster Bc may include more than one phage species. For each of the phage serogroups A, B, and F, common HindIII-restriction fragments of phage 3A (1700 bp), of 53 (4060 bp), and of 77 (8300 bp) were used for the preparation of probes specific to the phages of serogroups A, B, and F. These probes were very effective, making it possible to detect up to three different prophages in a given lysogenic strain at the same time. Restriction enzyme maps of phages 3A, 53, and 77, each representing a different serogroup, were constructed. The restriction maps of phage 3A and that of phage 77 are linear, whereas that of phage 53 is circular and exhibits a circular permutation. DNAs of the phages of serogroups A and F have cohesive ends. On each restriction map, the sites corresponding to specific probes are indicated. The size of intact genomic DNA of all phages estimated by PFGE varies within the range of 41.5-46.2 kb.


Asunto(s)
Tipificación de Bacteriófagos , ADN Viral/genética , Lisogenia , Provirus/genética , Fagos de Staphylococcus/genética , Análisis por Conglomerados , Sondas de ADN , ADN Circular/genética , ADN Circular/aislamiento & purificación , ADN Viral/aislamiento & purificación , Desoxirribonucleasas de Localización Especificada Tipo II , Provirus/clasificación , Provirus/aislamiento & purificación , Mapeo Restrictivo , Fagos de Staphylococcus/clasificación , Fagos de Staphylococcus/aislamiento & purificación
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