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1.
Article in English | MEDLINE | ID: mdl-22580855

ABSTRACT

BACKGROUND: The aim of the study was in vitro assessment and comparison of the antimicrobial activity of three types of silicone oils used in ophthalmic surgery. METHODS: The silicone oils (Arciolane 1300 centistokes, Arciolane 5500 centistokes and Oxane Hd, heavy silicone oil) were inoculated with microbes common in endophthalmitis and their growth was observed continuously. Control tests of microbial growth were performed on silicone oil-free media, i.e. saline and standard enrichment media. In both tested oils and control media, the microbes were cultured aerobically for 21 days, bacteria at 37 °C and yeasts and fungi at 30 °C. Prior to and during incubation at given intervals (days 0, 2, 4, 7, 9, 11, 14, 16, 18 and 21), 10 µl samples were taken from all test tubes. These were diluted in saline in a series of test tubes, with the minimum concentration reaching 10(-8). From each dilution, 25 µl were inoculated onto agar media. After 24 h of aerobic incubation at 37 °C (bacteria) and 48 h at 30 °C (yeasts and fungi), the grown colonies were counted and the numbers of colony-forming units in 1 ml (CFU/ml) were determined. RESULTS: In vitro, the highest antimicrobial effect was observed for the Oxane Hd silicone oil. CONCLUSIONS: If endophthalmitis is treated by pars plana vitrectomy, the application of Oxane Hd silicone oil into the vitreous cavity at the end of surgery may contribute to the elimination of microorganisms from the intraocular space but clinical trials are needed to assess its safety.


Subject(s)
Anti-Infective Agents/pharmacology , Bacteria/growth & development , Ophthalmologic Surgical Procedures/adverse effects , Silicone Oils , Aspergillus/growth & development , Candida/growth & development , Endophthalmitis/etiology , Endophthalmitis/microbiology , Humans , In Vitro Techniques , Silicone Oils/pharmacology
2.
Diagn Microbiol Infect Dis ; 71(3): 181-91, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21856106

ABSTRACT

A total of 78 bacterial strains with known ß-lactamases were used to optimize a rapid detection system consisting of multiplex polymerase chain reaction and melting curve analysis to amplify and identify blaTEM, blaSHV, and blaCTX-M genes in a single reaction. Additionally, to evaluate the applicability of this method, 32 clinical isolates of Escherichia coli displaying an extended-spectrum ß-lactamase phenotype from patients hospitalized at intensive care units were tested. Results were analyzed by the Rotor-Gene operating software and Rotor-Gene ScreenClust HRM Software. The individual melting curves differed by a temperature shift or curve shape, according to the presence of ß-lactamase genes. With the use of this method and direct sequencing, blaCTX-M-15-like was identified as the most prevalent ß-lactamase gene. In conclusion, this novel detection system seems to be a suitable tool for rapid detection of present ß-lactamase genes and their characterization.


Subject(s)
Escherichia coli/genetics , Multiplex Polymerase Chain Reaction , beta-Lactamases/genetics , Cross Infection/microbiology , Escherichia coli/isolation & purification , Escherichia coli Infections/microbiology , Humans , Intensive Care Units
3.
Klin Mikrobiol Infekc Lek ; 16(5): 164-70, 2010 Oct.
Article in Czech | MEDLINE | ID: mdl-21191874

ABSTRACT

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.


Subject(s)
Antifungal Agents/therapeutic use , Candidiasis, Invasive/drug therapy , Cross Infection/drug therapy , Intensive Care Units , Candidiasis, Invasive/diagnosis , Candidiasis, Invasive/etiology , Humans , Risk Factors
4.
Klin Mikrobiol Infekc Lek ; 16(1): 4-9, 2010 Feb.
Article in Czech | MEDLINE | ID: mdl-20401831

ABSTRACT

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.


Subject(s)
Antifungal Agents/pharmacology , Cattle/microbiology , Dogs/microbiology , Swine/microbiology , Yeasts/drug effects , Yeasts/isolation & purification , Animals , Candida/drug effects , Candida/isolation & purification , Microbial Sensitivity Tests
5.
Klin Mikrobiol Infekc Lek ; 15(4): 116-24, 2009 Aug.
Article in Czech | MEDLINE | ID: mdl-19937574

ABSTRACT

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.


Subject(s)
Candida/classification , Mycological Typing Techniques , Adolescent , Adult , Aged , Candida/isolation & purification , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Young Adult
6.
Klin Mikrobiol Infekc Lek ; 15(4): 125-30, 2009 Aug.
Article in Czech | MEDLINE | ID: mdl-19937575

ABSTRACT

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.


Subject(s)
Candida albicans/classification , Mycological Typing Techniques , Candida albicans/isolation & purification , Female , Humans , Male , Phenotype
7.
BMC Microbiol ; 9: 234, 2009 Nov 10.
Article in English | MEDLINE | ID: mdl-19903328

ABSTRACT

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.


Subject(s)
Candida/classification , Mycological Typing Techniques/methods , Random Amplified Polymorphic DNA Technique/methods , Algorithms , Automation/methods , Candida/genetics , Candida/isolation & purification , Cluster Analysis , DNA, Fungal/genetics , Genotype , Humans , Reproducibility of Results , Saccharomyces cerevisiae/classification , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/isolation & purification , Species Specificity
8.
Med Sci Monit ; 15(7): BR202-6, 2009 Jul.
Article in English | MEDLINE | ID: mdl-19564820

ABSTRACT

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.


Subject(s)
Enterobacteriaceae/enzymology , Enterobacteriaceae/isolation & purification , Residence Characteristics , beta-Lactamases/metabolism , Bacterial Typing Techniques , Czech Republic , Enterobacteriaceae/classification , Humans , Microbial Sensitivity Tests , Polymerase Chain Reaction , Prevalence
9.
New Microbiol ; 32(1): 67-76, 2009 Jan.
Article in English | MEDLINE | ID: mdl-19382671

ABSTRACT

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.


Subject(s)
Hematologic Neoplasms/complications , Klebsiella Infections , Klebsiella pneumoniae/isolation & purification , Pseudomonas Infections , Pseudomonas aeruginosa/isolation & purification , Anti-Bacterial Agents/pharmacology , Humans , Klebsiella Infections/epidemiology , Klebsiella Infections/microbiology , Klebsiella Infections/transmission , Klebsiella pneumoniae/classification , Klebsiella pneumoniae/genetics , Microbial Sensitivity Tests , Prevalence , Pseudomonas Infections/epidemiology , Pseudomonas Infections/microbiology , Pseudomonas Infections/transmission , Pseudomonas aeruginosa/classification , Pseudomonas aeruginosa/genetics
10.
Klin Mikrobiol Infekc Lek ; 14(5): 173-7, 2008 Oct.
Article in Slovak | MEDLINE | ID: mdl-19051165

ABSTRACT

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.


Subject(s)
Bacterial Proteins/metabolism , Klebsiella pneumoniae/isolation & purification , Leukemia/microbiology , beta-Lactamases/metabolism , Acute Disease , Humans , Klebsiella pneumoniae/enzymology
11.
Klin Mikrobiol Infekc Lek ; 14(3): 88-92, 2008 Jun.
Article in Czech | MEDLINE | ID: mdl-18688768

ABSTRACT

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.


Subject(s)
Antigens, Fungal/blood , Mycoses/diagnosis , beta-Glucans/blood , Humans , Immunocompromised Host , Proteoglycans , Sensitivity and Specificity , Serologic Tests
12.
New Microbiol ; 31(1): 97-104, 2008 Jan.
Article in English | MEDLINE | ID: mdl-18437847

ABSTRACT

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.


Subject(s)
Arthroplasty, Replacement/adverse effects , Culture Media , Polymerase Chain Reaction/methods , Prosthesis-Related Infections/diagnosis , Prosthesis-Related Infections/microbiology , Synovial Fluid/microbiology , Bacterial Infections/diagnosis , Bacterial Infections/microbiology , Bacteriological Techniques , DNA, Bacterial/analysis , Gram-Negative Bacteria/genetics , Gram-Negative Bacteria/isolation & purification , Gram-Positive Bacteria/genetics , Gram-Positive Bacteria/isolation & purification , Humans , Predictive Value of Tests , Sensitivity and Specificity
13.
Klin Mikrobiol Infekc Lek ; 13(5): 206-12, 2007 Oct.
Article in Slovak | MEDLINE | ID: mdl-17987570

ABSTRACT

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.


Subject(s)
Klebsiella pneumoniae/enzymology , beta-Lactam Resistance , beta-Lactamases/biosynthesis , Cross Infection/microbiology , Humans , Intensive Care Units , Klebsiella Infections/microbiology , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/isolation & purification , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , beta-Lactamases/genetics
14.
J Agric Food Chem ; 55(8): 3217-24, 2007 Apr 18.
Article in English | MEDLINE | ID: mdl-17381122

ABSTRACT

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.


Subject(s)
Antioxidants/analysis , Beverages/analysis , Food Preservation , Fruit/chemistry , Urine/chemistry , Vaccinium macrocarpon/chemistry , Adult , Bacterial Adhesion/drug effects , Double-Blind Method , Escherichia coli/drug effects , Female , Humans , Hydrogen-Ion Concentration , Oxidative Stress , Pilot Projects , Placebos , Urinary Tract Infections/prevention & control
15.
Klin Mikrobiol Infekc Lek ; 12(3): 117-123, 2006 Jun.
Article in Czech | MEDLINE | ID: mdl-17051473

ABSTRACT

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.


Subject(s)
Bacterial Infections/microbiology , Joint Prosthesis , Prosthesis-Related Infections/microbiology , Adult , Aged , Bacterial Infections/diagnosis , Bacteriological Techniques , DNA, Bacterial/analysis , Female , Humans , Male , Middle Aged , Polymerase Chain Reaction , Prosthesis-Related Infections/diagnosis , Sensitivity and Specificity
16.
FEMS Microbiol Lett ; 263(2): 163-8, 2006 Oct.
Article in English | MEDLINE | ID: mdl-16978351

ABSTRACT

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.


Subject(s)
Bacterial Typing Techniques , Staphylococcal Infections/microbiology , Staphylococcus/classification , Staphylococcus/isolation & purification , DNA Fingerprinting , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Humans , Novobiocin , Phylogeny , RNA, Ribosomal, 16S/analysis , RNA, Ribosomal, 16S/genetics , Ribotyping , Sequence Analysis, DNA , Staphylococcus/genetics , Staphylococcus/physiology
17.
New Microbiol ; 29(2): 121-5, 2006 Apr.
Article in English | MEDLINE | ID: mdl-16841552

ABSTRACT

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.


Subject(s)
Cross Infection/microbiology , Enterococcus faecalis/drug effects , Enterococcus faecium/drug effects , Gram-Positive Bacterial Infections/microbiology , Vancomycin Resistance , Cross Infection/epidemiology , Czech Republic/epidemiology , Enterococcus faecalis/isolation & purification , Enterococcus faecium/isolation & purification , Gram-Positive Bacterial Infections/epidemiology , Humans , Microbial Sensitivity Tests , Prevalence , Residence Characteristics
18.
Int J Syst Evol Microbiol ; 55(Pt 5): 1953-1958, 2005 Sep.
Article in English | MEDLINE | ID: mdl-16166694

ABSTRACT

Eight coagulase-negative, oxidase-negative and novobiocin-susceptible staphylococcal strains were isolated from the gastrointestinal tracts of South American squirrel monkeys (Saimiri sciureus L.). These strains were differentiated from known staphylococcal species on the basis of 16S rRNA gene and hsp60 gene sequencing, and from the most closely related species by using DNA-DNA hybridization, ribotyping, whole-cell protein profiles and biotyping. Phylogenetic analysis based on 16S rRNA gene sequences confirmed that these strains are members of the Staphylococcus aureus species group (99% similarity) but are biochemically similar to Staphylococcus piscifermentans, from which they can be phenotypically distinguished by resistance to polymyxin B, acid production from D-mannitol, the inability to hydrolyse aesculin and DNA and the absence of alpha-glucosidase. On the basis of these analyses, a novel species of the genus Staphylococcus is described, for which the name Staphylococcus simiae sp. nov. is proposed, with CCM 7213(T) (=LMG 22723(T)) as the type strain.


Subject(s)
Gastrointestinal Tract/microbiology , Saimiri/microbiology , Staphylococcus/classification , Animals , Bacterial Proteins/analysis , Bacterial Typing Techniques , Chaperonin 60/genetics , DNA, Bacterial/analysis , Genes, rRNA , Molecular Sequence Data , Nucleic Acid Hybridization , Phylogeny , RNA, Ribosomal, 16S/genetics , Ribotyping , Sequence Analysis, DNA , South America , Staphylococcus/genetics , Staphylococcus/isolation & purification , Staphylococcus/physiology
19.
J Med Microbiol ; 54(Pt 10): 965-967, 2005 Oct.
Article in English | MEDLINE | ID: mdl-16157551

ABSTRACT

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.


Subject(s)
Carrier State/epidemiology , Enterococcus/drug effects , Enterococcus/isolation & purification , Gram-Positive Bacterial Infections/epidemiology , Poultry/microbiology , Vancomycin Resistance/genetics , Animals , Bacterial Proteins/genetics , Carbon-Oxygen Ligases/genetics , Cloaca/microbiology , Cluster Analysis , Czech Republic/epidemiology , DNA Fingerprinting , DNA, Bacterial/genetics , Deoxyribonucleases, Type II Site-Specific , Enterococcus/classification , Enterococcus/genetics , Genetic Variation , Gram-Positive Bacterial Infections/microbiology , Humans , Peptide Synthases/genetics , Polymorphism, Restriction Fragment Length , Rectum/microbiology
20.
Klin Mikrobiol Infekc Lek ; 11(2): 47-50, 2005 Apr.
Article in Czech, English | MEDLINE | ID: mdl-15926119

ABSTRACT

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.


Subject(s)
Vancomycin Resistance , Vancomycin-Resistant Enterococci , Anti-Bacterial Agents/pharmacology , Enterococcus , Humans , Polymerase Chain Reaction , Prevalence
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