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1.
Klin Mikrobiol Infekc Lek ; 16(1): 4-9, 2010 Feb.
Artigo em Tcheco | MEDLINE | ID: mdl-20401831

RESUMO

Yeasts frequently colonize various kinds of domestic animals, but may also cause serious diseases. The aim of this study was to identify yeast isolates collected from dogs, cows and pigs, and to determine their in vitro antifungal susceptibility. Fifty-six yeast isolates from dogs (n = 24), cows (n = 20), and pigs (n = 12) were investigated. Appearance of colonies grown on Sabouraud agar, micromorphology on rice agar, as well as assimilation and fermentation of various carbon and nitrogen sources were evaluated. Susceptibility to six antifungals (flucytosine, amphotericin B, miconazole, ketoconazole, itraconazole and fluconazole) was determined semiquantitatively using the commercially available Fungitest kit (Bio-Rad Laboratories). Ten yeast species were identified in dogs with relatively even distribution. On the other hand, cow and pig were clearly dominated by Candida krusei (from 7 species) and Candida rugosa (from 5 species), respectively. Further, most of yeast isolates exhibited good susceptibility to the antifungals tested particularly to amphotericin B, ketoconazole and itraconazole. Based on results, it can be concluded that significant differences in the species spectrum and distribution were documented between groups of yeasts from dogs, cows and pigs. This is probably due to different environmental conditions and the endogenous origin of the yeast isolates. Mostly good susceptibility to systemic antifungals should positively influence the therapy of diseases caused by yeasts in veterinary medicine.


Assuntos
Antifúngicos/farmacologia , Bovinos/microbiologia , Cães/microbiologia , Suínos/microbiologia , Leveduras/efeitos dos fármacos , Leveduras/isolamento & purificação , Animais , Candida/efeitos dos fármacos , Candida/isolamento & purificação , Testes de Sensibilidade Microbiana
2.
Klin Mikrobiol Infekc Lek ; 16(5): 164-70, 2010 Oct.
Artigo em Tcheco | MEDLINE | ID: mdl-21191874

RESUMO

Candidemia and invasive candidiasis are the most frequent mycoses in critically ill patients in intensive care units. Recently, the number of systemic antifungal agents has increased, leading to improved treatment options. Yet these infections remain to be characterized by poor prognosis and high mortality rates. The most important predisposing factors are yeast colonization of the mucosa or skin and damage to the integrity of the host's natural barriers. Early diagnosis of invasive candidiasis is difficult, since its clinical manifestations are not characteristic and the laboratory techniques are time-consuming and not completely reliable. The currently available treatments comprise three groups of antifungals: triazoles, polyenes and echinocandins. For its effectiveness, low toxicity and reasonable price, fluconazole is the most widespread drug currently used to treat systemic yeast infections. However, despite high treatment costs, echinocandins are becoming the drug of choice. The advantages are a broad spectrum of species, safe administration to patients with kidney and liver damage, minimal drug interactions and fungicidal effects. Candidemia may often be positively influenced by replacing an intravenous catheter. Despite earlier controversy, the latest treatment strategies clearly recommend its removal. Although antifungal prophylaxis lowers the incidence of invasive candidiasis, it is considered to be useful only if targeted to high-risk groups of patients. Empirical treatment of febrile patients not responding to broad-spectrum antibiotic therapy is only effective in wards with a higher incidence of systemic candidiasis, in patients with risk factors and if other causes are reliably excluded.


Assuntos
Antifúngicos/uso terapêutico , Candidíase Invasiva/tratamento farmacológico , Infecção Hospitalar/tratamento farmacológico , Unidades de Terapia Intensiva , Candidíase Invasiva/diagnóstico , Candidíase Invasiva/etiologia , Humanos , Fatores de Risco
3.
BMC Microbiol ; 9: 234, 2009 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-19903328

RESUMO

BACKGROUND: Rapid, easy, economical and accurate species identification of yeasts isolated from clinical samples remains an important challenge for routine microbiological laboratories, because susceptibility to antifungal agents, probability to develop resistance and ability to cause disease vary in different species. To overcome the drawbacks of the currently available techniques we have recently proposed an innovative approach to yeast species identification based on RAPD genotyping and termed McRAPD (Melting curve of RAPD). Here we have evaluated its performance on a broader spectrum of clinically relevant yeast species and also examined the potential of automated and semi-automated interpretation of McRAPD data for yeast species identification. RESULTS: A simple fully automated algorithm based on normalized melting data identified 80% of the isolates correctly. When this algorithm was supplemented by semi-automated matching of decisive peaks in first derivative plots, 87% of the isolates were identified correctly. However, a computer-aided visual matching of derivative plots showed the best performance with average 98.3% of the accurately identified isolates, almost matching the 99.4% performance of traditional RAPD fingerprinting. CONCLUSION: Since McRAPD technique omits gel electrophoresis and can be performed in a rapid, economical and convenient way, we believe that it can find its place in routine identification of medically important yeasts in advanced diagnostic laboratories that are able to adopt this technique. It can also serve as a broad-range high-throughput technique for epidemiological surveillance.


Assuntos
Candida/classificação , Técnicas de Tipagem Micológica/métodos , Técnica de Amplificação ao Acaso de DNA Polimórfico/métodos , Algoritmos , Automação/métodos , Candida/genética , Candida/isolamento & purificação , Análise por Conglomerados , DNA Fúngico/genética , Genótipo , Humanos , Reprodutibilidade dos Testes , Saccharomyces cerevisiae/classificação , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/isolamento & purificação , Especificidade da Espécie
4.
Med Sci Monit ; 15(7): BR202-6, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19564820

RESUMO

BACKGROUND: Problematic bacteria in the community include enterobacteria which produce extended-spectrum beta-lactamases. As yet there is no description of the prevalence of these bacteria in persons in the community in the Czech Republic. Therefore the main goal of this study was to determine the prevalence of ESBL-positive enterobacteria in the gastrointestinal tracts of subjects in the community in the Czech Republic. MATERIAL/METHODS: Rectal swabs from the investigated subjects were inoculated onto chromID ESBL selective medium and enterobacteria were identified by the Vitek2 automated system. ESBL were detected using a modified DDST test. The results were confirmed by PCR and direct sequencing of CTX-M-positive amplicons. RESULTS: A total of 579 rectal swabs from subjects in the community were analyzed and ESBL production was both phenotypically and genotypically confirmed in 7 isolates. Thus the prevalence of ESBL-positive bacteria in the gastrointestinal tracts of the persons in the community was 1.2%. All the cases were Escherichia coli strains producing the CTX-M-type ESBL. CTX-M-15 was the most prevalent type in this group of isolates. CONCLUSIONS: The presented results are in accord with other authors' studies and suggest that the epidemiologic profile of ESBL-positive enterobacteria in the Czech Republic is comparable to that in other European countries.


Assuntos
Enterobacteriaceae/enzimologia , Enterobacteriaceae/isolamento & purificação , Características de Residência , beta-Lactamases/metabolismo , Técnicas de Tipagem Bacteriana , República Tcheca , Enterobacteriaceae/classificação , Humanos , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Prevalência
5.
New Microbiol ; 32(1): 67-76, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19382671

RESUMO

The aim of the study was to determine the prevalence of Pseudomonas aeruginosa and Klebsiella pneumoniae strains in patients with acute leukemias, to assess their clinical significance, and to define the sources and ways of their spread using genetic analysis. Thirty-four patients were investigated during the observed period. Twenty-one strains of Pseudomonas aeruginosa and 35 strains of Klebsiella pneumoniae were isolated from patient samples. In the case of Pseudomonas aeruginosa, 47.6% of strains were identified as pathogens and caused infection. By contrast, only 4 isolates (11.4%) of Klebsiella pneumoniae could be regarded as etiological agents of bacterial infection. Based on the obtained results, Klebsiella pneumoniae strains are assumed to be of mostly endogenous origin. In the case of Pseudomonas aeruginosa strains, the proportion of identical strains detected in various patients was higher and exogenous sources were more significant. In addition, our results confirmed the ability of Pseudomonas aeruginosa strains to survive on a particular site in the hospital for a longer time.


Assuntos
Neoplasias Hematológicas/complicações , Infecções por Klebsiella , Klebsiella pneumoniae/isolamento & purificação , Infecções por Pseudomonas , Pseudomonas aeruginosa/isolamento & purificação , Antibacterianos/farmacologia , Humanos , Infecções por Klebsiella/epidemiologia , Infecções por Klebsiella/microbiologia , Infecções por Klebsiella/transmissão , Klebsiella pneumoniae/classificação , Klebsiella pneumoniae/genética , Testes de Sensibilidade Microbiana , Prevalência , Infecções por Pseudomonas/epidemiologia , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/transmissão , Pseudomonas aeruginosa/classificação , Pseudomonas aeruginosa/genética
6.
Klin Mikrobiol Infekc Lek ; 15(4): 116-24, 2009 Aug.
Artigo em Tcheco | MEDLINE | ID: mdl-19937574

RESUMO

BACKGROUND: The biotyping system according to Odds and Abbott belongs to the most frequently used phenotypic methods. The aim of the study was to evaluate its discriminatory power in our laboratory. In addition, biotypes of isolates obtained from various body locations, present in defined age groups of patients and in males and females were also compared. MATERIAL AND METHODS: A total of 343 clinical isolates were typed belonging to six most frequent Candida spp. Nine types of tests for biotyping were prepared: sorbose, citrate and urea assimilation, tolerance to pH 1.4, pH 1.55 and higher concentration of NaCl, resistance to sodium periodate and boric acid and the ability to grow on MacConkey agar. RESULTS: Forty-one biotypes were found among 230 C. albicans isolates, nine among 21 C. glabrata, 13 among 25 C. parapsilosis, 12 among 25 C. krusei and five biotypes among 18 C. lusitaniae isolates. Contrary to other species, all of 18 C. tropicalis isolates belonged to the same biotype. In accordance with previously published reports, high discriminatory power of the method was found with Simpson's diversity index for C. albicans reaching 0.92. On the other hand, reproducibility was relatively low; from 12 randomly chosen C. albicans isolates tested repeatedly, only two showed identical results, five differed in one test and the others in several tests. Analysis of the occurrence of individual biotypes related to different anatomical locations, age groups and sexes of patients revealed neither statistically significant variations in distribution nor predilection of any single biotype. These findings suggest that the source of infection was endogenous in most cases. In comparison with results of similar studies, marked discrepancies in profiles of predominant biotypes were found, probably due to slight differences in composition of the test media or distinctive evaluation of results; however, it may reflect also geographical specificity of isolates. CONCLUSION: It can be concluded that the main advantages of the Odds biotyping system are high discriminatory power and cost-effectiveness. On the other hand, discrepancies in reproducibility of results as well as relatively long period for preparation of test media and for achieving of results decline its usefulness for epidemiological studies.


Assuntos
Candida/classificação , Técnicas de Tipagem Micológica , Adolescente , Adulto , Idoso , Candida/isolamento & purificação , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Adulto Jovem
7.
Klin Mikrobiol Infekc Lek ; 15(4): 125-30, 2009 Aug.
Artigo em Tcheco | MEDLINE | ID: mdl-19937575

RESUMO

BACKGROUND: Morphotyping is a phenotypic method for strain differentiation of yeast cultures based on the comparison of appearance of their surfaces and fringes. For its simplicity and cost-effectiveness, it is recommended as an alternative tool to genotyping of Candida albicans. Our study aimed at verifying its usefulness for typing a group of C. albicans clinical isolates with emphasis on discrimination power. Prevalence of specific morphotypes in body locations with more tested isolates and according to age groups and gender of patients was also evaluated. MATERIAL AND METHODS: The tested group comprised 97 C. albicans isolates from 93 patients. With a cotton swab, 24-hour cultures were streaked onto malt extract agar. The surface and fringe morphology was assessed after 10-day incubation at 30 degrees C. RESULTS: A total of 30 morphotypes were detected. The most frequent one had a code profile 7240 (n = 13, 13.4 %), followed by 5240 (n = 11) and 7340 (n = 9). The discrimination power calculated by Simpson's index of diversity reached 0.95. Results of reproducibility testing were identical in 54.6 % of isolates, another 36.1 % of isolates differed in one character. Some isolates (n = 35) were biotyped by our modification of Odds method. Using morphological criteria, 14 different phenotypes were defined, as opposed to 8 found by biotyping. Statistically significant differences were found when comparing morphotypes from the oropharynx, vagina and urine; the same was true for male and female morphotypes. CONCLUSION: Our results clearly documented very good discrimination power of morphotyping for C. albicans isolates. This advantage as well as easiness to perform and low costs but markedly lower reproducibility in comparison with molecular genetic techniques makes it an optimal typing method for first-line use. Evaluation of morphological diversity of strains by this method can be further utilized in the studies of virulence, switching phenomenon or antifungal resistance.


Assuntos
Candida albicans/classificação , Técnicas de Tipagem Micológica , Candida albicans/isolamento & purificação , Feminino , Humanos , Masculino , Fenótipo
8.
New Microbiol ; 31(1): 97-104, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18437847

RESUMO

This prospective study compared PCR and culture techniques in the diagnosis of prosthetic joint infection (PJI). We obtained joint fluid samples (JFS; n=115) from patients who had failed total joint arthroplasty between January 2003 and June 2005; 49 were positive for PJI according to established strict criteria. JFS were analyzed by PCR (n=35; control n=66) or culture (n=46, control n=48). PCR was positive in 71% of PJI cases, resulting in sensitivity, specificity, accuracy, positive predictive value, negative predictive value, and likelihood ratio for positive results as follows: 0.71; 0.97; 0.88; 0.93; 0.87 and 23.6, respectively. Culture was positive in 44% of PJI samples. Corresponding statistics were 0.44; 0.94; 0.69; 0.87; 0.63 and 7.0, respectively. Significantly higher sensitivity, accuracy and negative predictive values were calculated for PCR versus culture, and there was 83% concordance between the results of intraoperative culture and PCR detection of causative bacteria. Therefore, we conclude that PCR analysis of synovial fluid increases the utility of pre-operative aspiration for patients who require revision total joint surgery.


Assuntos
Artroplastia de Substituição/efeitos adversos , Meios de Cultura , Reação em Cadeia da Polimerase/métodos , Infecções Relacionadas à Prótese/diagnóstico , Infecções Relacionadas à Prótese/microbiologia , Líquido Sinovial/microbiologia , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/microbiologia , Técnicas Bacteriológicas , DNA Bacteriano/análise , Bactérias Gram-Negativas/genética , Bactérias Gram-Negativas/isolamento & purificação , Bactérias Gram-Positivas/genética , Bactérias Gram-Positivas/isolamento & purificação , Humanos , Valor Preditivo dos Testes , Sensibilidade e Especificidade
9.
Klin Mikrobiol Infekc Lek ; 14(3): 88-92, 2008 Jun.
Artigo em Tcheco | MEDLINE | ID: mdl-18688768

RESUMO

The incidence of deep fungal infections has increased in immunocompromised patients. Prompt and reliable diagnosis is crucial for improving the outcome of this life-threatening disease. Non-culture-based diagnostic methods, such as antigen (antibody) detection, are very often used. Detection of 1,3-Beta-D-glucan, a panfungal antigen from fungal cell walls, is one of the new commercially available diagnostic techniques that appear to be useful for patients with suspicion of fungal infection (aspergillosis, candidiasis and infection with rare species). This test can be used directly for early diagnosis of invasive fungal infection or as a tool to identify false positive results of other methods.


Assuntos
Antígenos de Fungos/sangue , Micoses/diagnóstico , beta-Glucanas/sangue , Humanos , Hospedeiro Imunocomprometido , Proteoglicanas , Sensibilidade e Especificidade , Testes Sorológicos
10.
Klin Mikrobiol Infekc Lek ; 14(5): 173-7, 2008 Oct.
Artigo em Sk | MEDLINE | ID: mdl-19051165

RESUMO

BACKGROUND: Currently, important bacterial beta-lactamases of increasing clinical significance include AmpC enzymes. The aim was to assess their occurrence in Klebsiella pneumoniae strains isolated from patients with haematological malignancies in a prospective study. MATERIAL AND METHODS: Over a 2-month period, strains of the species were isolated from clinical material obtained from patients hospitalized at the Department of Haemato-Oncology of the University Hospital Olomouc. The strains were identified using standard microbiological techniques and the Vitek 2 automated system. Production of AmpC beta-lactamases was roughly determined by phenotypic tests and subsequently confirmed by PCR detection of genes encoding these enzymes. RESULTS: During the above-mentioned period, a total of 35 K. pneumoniae isolates were collected. In 7 of them, production of AmpC beta-lactamases was preliminarily detected by phenotypic test. The multiplex PCR method confirmed phenotyping and determined DHA types in all the isolates. CONCLUSIONS: All AmpC-positive isolates were false-susceptible to at least one of the tested third-generation cephalosporins. In one patient, clinically apparent infection caused by this strain was documented. The reported results suggest the possibility of occurrence of AmpC-beta-lactamases in K. pneumoniae strains with clinical significance.


Assuntos
Proteínas de Bactérias/metabolismo , Klebsiella pneumoniae/isolamento & purificação , Leucemia/microbiologia , beta-Lactamases/metabolismo , Doença Aguda , Humanos , Klebsiella pneumoniae/enzimologia
11.
J Agric Food Chem ; 55(8): 3217-24, 2007 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-17381122

RESUMO

This study assessed the effect of an 8 week consumption of dried cranberry juice (DCJ) on 65 healthy young women. Basic biochemical and hematological parameters, antioxidant status, presence of metabolites in urine, and urine ex vivo antiadherence activity were determined throughout the trial. A 400 mg amount of DCJ/day had no influence on any parameter tested. A 1200 mg amount of DCJ/day resulted in a statistically significant decrease in serum levels of advanced oxidation protein products. This specific protective effect against oxidative damage of proteins is described here for the first time. Urine samples had an inhibitory effect on the adhesion of uropathogenic Escherichia coli strains, but no increase in urine acidity was noted. Hippuric acid, isomers of salicyluric and dihydroxybenzoic acids, and quercetin glucuronide were identified as the main metabolites. In conclusion, cranberry fruits are effective not only in the prevention of urinary tract infection but also for the prevention of oxidative stress.


Assuntos
Antioxidantes/análise , Bebidas/análise , Conservação de Alimentos , Frutas/química , Urina/química , Vaccinium macrocarpon/química , Adulto , Aderência Bacteriana/efeitos dos fármacos , Método Duplo-Cego , Escherichia coli/efeitos dos fármacos , Feminino , Humanos , Concentração de Íons de Hidrogênio , Estresse Oxidativo , Projetos Piloto , Placebos , Infecções Urinárias/prevenção & controle
12.
Klin Mikrobiol Infekc Lek ; 13(5): 206-12, 2007 Oct.
Artigo em Sk | MEDLINE | ID: mdl-17987570

RESUMO

OBJECTIVES: The study aimed at the assessment of the prevalence of ESBL-positive isolates of Klebsiella pneumoniae in intensive care patients and their molecular biology analysis. MATERIAL AND METHODS: Over a 5-month period, Klebsiella pneumoniae strains were isolated from patients hospitalized at the Department of Anaesthesiology and Resuscitation of the University Hospital in Olomouc. For each isolate, an antibiogram was performed by the standard microdilution method and the production of ESBL was determined by the modified double-disk synergy test. PCR was used to demonstrate the presence of the blaTEM and blaSHV genes. The isolates producing SHV- and TEM-types of beta-lactamases were typed using the restriction fragment length polymorphism (RFLP) method to identify the most common mutations responsible for the development of an ESBL phenotype. Similar or identical isolates were determined by pulsed-field gel electrophoresis (PFGE) of DNA fragments cleaved by the XbaI restriction endonuclease. RESULTS: A total of 67 isolates of Klebsiella pneumoniae were obtained. In 13 of them, the production of ESBL was detected and the presence of the blaSHV gene was confirmed by PCR. Restriction cleavage by NheI revealed mutations at position 238 in all SHV-positive PCR products. The restriction analysis did not confirm the presence of the gene encoding TEM-type extended-spectrum beta-lactamase. Molecular biology typing by PFGE detected the presence of 11 different strains. CONCLUSIONS: In the observed group of intensive care patients, the prevalence of ESBL-positive strains of Klebsiella pneumoniae reached 19.4 %. The analysis of SHV and TEM products of PCR by the RFLP method showed the prevalence of SHV-type ESBL. Overall, 84.6 % of the strains had unique restriction profiles. The results suggest both high levels of hygienic and epidemiological measures at the monitored department and rational antibiotic policy.


Assuntos
Klebsiella pneumoniae/enzimologia , Resistência beta-Lactâmica , beta-Lactamases/biossíntese , Infecção Hospitalar/microbiologia , Humanos , Unidades de Terapia Intensiva , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/isolamento & purificação , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , beta-Lactamases/genética
13.
FEMS Microbiol Lett ; 263(2): 163-8, 2006 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16978351

RESUMO

Five isolates of coagulase-negative staphylococci were obtained from human urine, the gastrointestinal tract of squirrel monkeys, pig skin and from the environment. All key biochemical characteristics of the tested strains corresponded with the description of Staphylococcus xylosus species. However, partial 16S rRNA gene sequences obtained from analysed strains corresponded with those of Staphylococcus nepalensis reference strains, except for two strains which differed in one residue. Ribotyping with EcoRI and HindIII restriction enzymes, whole cell protein profile analysis performed by SDS-PAGE and SmaI macrorestriction analysis were used for more precise characterization and identification of the analysed strains. Obtained results showed that EcoRI and HindIII ribotyping and whole cell protein fingerprinting are suitable and reliable methods for the differentiation of S. nepalensis strains from the other novobiocin resistant staphylococci, whereas macrorestriction analysis was found to be a good tool for strain typing. The isolation of S. nepalensis is sporadic, and according to our best knowledge this study is the first report of the occurrence of this species in human clinical material as well as in other sources.


Assuntos
Técnicas de Tipagem Bacteriana , Infecções Estafilocócicas/microbiologia , Staphylococcus/classificação , Staphylococcus/isolamento & purificação , Impressões Digitais de DNA , DNA Bacteriano/química , DNA Bacteriano/genética , Humanos , Novobiocina , Filogenia , RNA Ribossômico 16S/análise , RNA Ribossômico 16S/genética , Ribotipagem , Análise de Sequência de DNA , Staphylococcus/genética , Staphylococcus/fisiologia
14.
New Microbiol ; 29(2): 121-5, 2006 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-16841552

RESUMO

Between July 1, 2002 and December 31, 2003, rectal swabs from both hospitalized patients and community subjects in the Czech Republic were taken to ascertain the prevalence of vancomycin-resistant enterococci (VRE). The swabs were used for isolating and identifying enterococci and their susceptibility to antibiotics. Vancomycin resistance phenotypes were verified by PCR detection of vanA, vanB, vanC1 and vanC2 genes. A molecular biology analysis was performed in Enterococcus faecium VanA strains. During the observed period, 2691 rectal swabs from the hospitalized patients and 6529 rectal swabs from the subjects in community setting were examined. In total, 31 VRE of hospital origin and 13 community-population strains were isolated. The prevalence of VRE in the gastrointestinal tract was 1.9% in the hospitalized patients and 0.4% in the community subjects. The prevailing strains were Enterococcus faecium VanA (61.3%) in the VRE of hospital origin and Enterococcus gallinarum VanC (46.2%) in the community VRE. Mutual comparison between the hospital and community Enterococcus faecium VanA strains showed no similarity.


Assuntos
Infecção Hospitalar/microbiologia , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecium/efeitos dos fármacos , Infecções por Bactérias Gram-Positivas/microbiologia , Resistência a Vancomicina , Infecção Hospitalar/epidemiologia , República Tcheca/epidemiologia , Enterococcus faecalis/isolamento & purificação , Enterococcus faecium/isolamento & purificação , Infecções por Bactérias Gram-Positivas/epidemiologia , Humanos , Testes de Sensibilidade Microbiana , Prevalência , Características de Residência
15.
Klin Mikrobiol Infekc Lek ; 12(3): 117-123, 2006 Jun.
Artigo em Tcheco | MEDLINE | ID: mdl-17051473

RESUMO

BACKGROUND: Successful treatment of patients with periprosthetic infection (PPI) requires a correct diagnosis supported by clinical, imaging, microbiological and other laboratory evidence. Equally important is to determine the causative agent of the infection as this may affect the subsequent treatment strategy. The paper aims at presenting cultivation results in a group of PPI patients and their comparison with molecular biology methods. MATERIALS AND METHODS: Material obtained during operations of PPI patients was examined by both the standard culture methods and the PCR technique to identify the etiological agents. The results were statistically compared. RESULTS: In total, the group comprised 49 patients with hip, knee or elbow replacement infection verified during the operation. In 42 cases (85.7 %), the infectious agent was identified by cultivation. The infection was most frequently (62.0 %) caused by coagulase-negative staphylococci and Staphylococcus aureus. Monomicrobial and polymicrobial infections were demonstrated in 35 (71.4 %) and 7 (14.3 %) patients, respectively. The PCR assay aimed at the 16S rRNA segment of bacterial DNA was performed in 35 patients, with positive results in 25 cases (71.4 %). In 82.6 %, the agents detected by specific PCR were consistent with the cultivation results. A statistically non-significant difference in sensitivity between the two methods was found, with higher specificity in the case of PCR. CONCLUSION: Standard cultivation methods remain a highly successful and useful tool for identifying the PPI causative agents.


Assuntos
Infecções Bacterianas/microbiologia , Prótese Articular , Infecções Relacionadas à Prótese/microbiologia , Adulto , Idoso , Infecções Bacterianas/diagnóstico , Técnicas Bacteriológicas , DNA Bacteriano/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Infecções Relacionadas à Prótese/diagnóstico , Sensibilidade e Especificidade
16.
J Med Microbiol ; 54(Pt 10): 965-967, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16157551

RESUMO

In the period between September 2002 and May 2004, a total of 6023 rectal swabs from humans in the Czech Republic were evaluated and 821 Enterococcus spp. strains were isolated. Nine strains (1.1 %) were identified as vancomycin-resistant enterococci (VRE). Two strains were VanA Enterococcus faecium, one strain was VanB Enterococcus faecalis and six strains were VanC Enterococcus casseliflavus. In total, 527 Enterococcus spp. strains were isolated from poultry breeds of which 11 (2.1 %) were VRE. Most (54.5 %) were identified as VanA E. faecium. Cluster analysis of SmaI-generated macrorestriction patterns showed high variability in both human and animal VRE strains and no relatedness between strains from the two sources.


Assuntos
Portador Sadio/epidemiologia , Enterococcus/efeitos dos fármacos , Enterococcus/isolamento & purificação , Infecções por Bactérias Gram-Positivas/epidemiologia , Aves Domésticas/microbiologia , Resistência a Vancomicina/genética , Animais , Proteínas de Bactérias/genética , Carbono-Oxigênio Ligases/genética , Cloaca/microbiologia , Análise por Conglomerados , República Tcheca/epidemiologia , Impressões Digitais de DNA , DNA Bacteriano/genética , Desoxirribonucleases de Sítio Específico do Tipo II , Enterococcus/classificação , Enterococcus/genética , Variação Genética , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Peptídeo Sintases/genética , Polimorfismo de Fragmento de Restrição , Reto/microbiologia
17.
Artigo em Inglês | MEDLINE | ID: mdl-16601773

RESUMO

The diagnosis of low grade prosthetic joint infection is difficult and time consuming. Nested-PCR for universal bacterial DNA segments detection of "orthopaedic" bacteria was tested in a laboratory setting. This method is based on amplification of the 16S bacterial ribosomal RNA coding sequences. 11 species of the most frequent bacterial pathogens (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus agalactiae, Enterococcus faecium, Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Serratia marcescens) involved in prosthetic joint infections were studied. All could be detected rapidly and sensitively by this method.


Assuntos
Bactérias/isolamento & purificação , Infecções Bacterianas/microbiologia , Prótese Articular , Infecções Relacionadas à Prótese/microbiologia , Primers do DNA , Humanos , Articulações/microbiologia , Reação em Cadeia da Polimerase
18.
Klin Mikrobiol Infekc Lek ; 11(1): 20-4, 2005 Feb.
Artigo em Tcheco | MEDLINE | ID: mdl-15832259

RESUMO

BACKGROUND: One of the problems of contemporary medicine is an increasing number of bacterial strains with hazardous phenotypes of resistance. This is also true for neonatal units where nosocomial infections caused by multiresistant bacteria pose a serious threat to newborns. The feared bacterial pathogens include Klebsiella pneumoniae strains producing AmpA Extended-Spectrum Beta-Lactamases. The study focused on the molecular biology characteristics of ESBL-positive strains of K. pneumoniae collected in the Neonatal Unit of the Teaching Hospital in Olomouc (THO). MATERIALS AND METHODS: Clinical material from newborns hospitalized in the THO Neonatal Unit between January and June 2004 was used to isolate and determine K. pneumoniae strains by standard identification procedures. Their susceptibility to antibiotics was tested using a dilution micromethod. A Double-Disk Synergy Test was used for phenotype determination of ESBL production. The bla gene coding ESBL production was demonstrated by PCR. Molecular biology characteristics of ESBL-positive strains utilized the genomic DNA isolation, XbaI restrictase digestion and PFGE differentiation. The acquired restriction maps of individual isolates were compared using the GelCompare software and their relationship was determined. The selection pressure of antimicrobial agents was assessed according to the absolute number of defined daily doses of individual antibiotics. RESULTS: During the monitored period, 112 K. pneumoniae strains were isolated in total. In 22 of them (19.6%), the TEM-type ESBL production was determined. ESBL-positive strains were only observed in upper respiratory tract and rectal swabs collected from newborns with no signs of infection. The molecular biology analysis showed that 21 ESBL-positive strains had an identical restriction profile, i.e. they were very likely to be identical. The selection pressure of third- and fourth-generation cephalosporins was very low over the observed period and their consumption accounted for 1.9 % of all administered antimicrobial agents. CONCLUSION: The results presented above suggest that ESBL-positive strains of K. pneumoniae occurred in the THO Neonatal Unit due to clonal and horizontal spread from an unidentified source.


Assuntos
Infecção Hospitalar/microbiologia , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/genética , beta-Lactamases/metabolismo , Farmacorresistência Bacteriana/genética , Humanos , Recém-Nascido , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/isolamento & purificação , Testes de Sensibilidade Microbiana , Berçários Hospitalares , Reação em Cadeia da Polimerase , Resistência beta-Lactâmica , beta-Lactamases/genética
19.
Klin Mikrobiol Infekc Lek ; 11(2): 47-50, 2005 Apr.
Artigo em Tcheco, Inglês | MEDLINE | ID: mdl-15926119

RESUMO

AIM OF THE STUDY: The presented study aimed at determining the prevalence of vancomycin-resistant enterococci (VRE) in rectal swabs taken from both patients in the Teaching Hospital in Olomouc (THO), Czech Republic, and subjects from the community setting of the hospital's catchment area. MATERIALS AND METHODS: Between July 1, 2002 and July 1, 2003, rectal swabs were taken from the THO patients as well as individuals from the community catchment area to be utilized for isolating and identifying enterococci and their suscetibility to antibiotics. Vancomycin resistance phenotypes were verified by PCR detection of vanA, vanB, vanC1 and vanC2 genes. A molecular biology analysis was performed in VanA Enterococcus faecium strains. To determine the relationship of strains, macrorestriction analysis of the total chromosomal DNA digested with SmaI restriction endonuclease was used. RESULTS: During the observed period, 2,157 rectal swabs from the hospitalized patients and 4,874 rectal swabs from the subjects in community setting were examined. In total, 27 VRE of hospital origin and 13 community-population strains were isolated. The prevalence of VRE in the gastrointestinal tract was 2.3 % in the hospitalized patients and 0.6 % in the community subjects. The prevailing strains were Enterococcus faecium VanA (70.4 %) in the VRE of hospital origin and Enterococcus gallinarum VanC (46.2 %) in the community VRE. Mutual comparison between the hospital and community VRE showed no similarity. CONCLUSION: In the Czech Republic, VRE were proved both in community and hospital settings. Their prevalence in rectal swabs is low and does not exceed the values reported in other European countries. The author describes the chief characteristics of the most important quinolone antibiotics, including preparations either in their development stage or whose development has been prematurely interrupted because of adverse side-effects. The list includes all preparations that are or were temporarily registered in the Czech Republic.


Assuntos
Resistência a Vancomicina , Enterococos Resistentes à Vancomicina , Antibacterianos/farmacologia , Enterococcus , Humanos , Reação em Cadeia da Polimerase , Prevalência
20.
Artigo em Inglês | MEDLINE | ID: mdl-12572892

RESUMO

There was followed the development of serological findings in patients with proved positivity only in classes IgA and/or IgM of chlamydial antibodies (without IgG), which can be suspected of showing "false" positivity. 184 patients were repeatedly examined for chlamydial antibodies in their sera (interval between collections up to three months) using a genus specific rELISA. Sera were also tested for the evidence of IgM antibodies against capside antigen of Epstein-Barr virus (EBV) and against cytomegalovirus (CMV) using ELISA methods. In 75 (40.8%) of patients, IgA/IgM individual positivities were demonstrated even during the following sample test(s). In 28 (15.2%) of them, IgG evidence preceded and in 29 (15.7%) other patients positive seroconversion followed in this class. In 13 (7.1%) patients, IgG antibodies disappeared and subsequently reappeared. Only in 39 (21.2%) of these probands, antibodies IgA/IgM were not demonstrated at another examination. Active EBV, resp. CMV infection was proved in 24 (13.0%), resp. in 18 (9.8%) of patients. It is concluded that the evidence of positivities only in classes IgA and/or IgM mostly signal the onset of a primary infection (reinfection) or an active infection in patients with IgG production failures respectively. In these cases, a "false" positivity can be supposed to occur only in a minor extent.


Assuntos
Anticorpos Antibacterianos/sangue , Infecções por Chlamydia/diagnóstico , Imunoglobulina A/sangue , Imunoglobulina M/sangue , Adolescente , Adulto , Idoso , Anticorpos Antivirais/sangue , Criança , Pré-Escolar , Chlamydia/imunologia , Citomegalovirus/imunologia , Reações Falso-Positivas , Feminino , Herpesvirus Humano 4/imunologia , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Testes Sorológicos
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