Medycyna doswiadczalna i mikrobiologia最新文献

Introduction: Corynebacterium diphtheriae can cause various infections such as diphtheria, wound infections, septic arthritis, bacteraemia and endocarditis. Different virulence properties of the isolates might be related to different virulence factors expressed by the isolates. The objective of this study was to explore whether whole cell protein profiling might be useful in prediction of pathogenic properties of C. diphtheriae isolates.

Methods: C. disphtheriae isolates collected from diphtheria, invasive and local infections and from asymptomatic carriers in Poland, France, New Caledonia and Canada in 1950-2014 were investigated using whole cell protein profile analysis.

Results: All the examined isolates were divided into two clades: A and B with similarity about 47%, but clade B was represented by only one isolate. The clade A was divided in two subclades A.I NS .II with similarity 53,2% and then into four groups: A.Ia, A.Ib, A.Ic and A.Id. The comparative analysis did not distinguish clearly toxigenic and nontoxigenic isolates as well as invasive and noninvasive isolates.

Conclusions: Whole cell protein profile analysis of C. diphtheria exhibits good concordance with other genotyping methods but this method is not able to distinguish clearly invasive from non-invasive isolates.

Introduction: Verocytotoxin-producing E. coli (VTEC) are a significant cause of haemor- rhagic colitis (HC) and haemolytic uremic syndrome (HUS) in humans. Because VTEC isolates are usually present in patients' feces for only a limited period of time serodiagnosis based on the purified antigens have become the useful tool for laboratory diagnosis and monitoring of prevalence of VTEC infections. The aim of the present study was to evaluate the usefulness of in-house obtained recombinant proteins Tir, intymin and verocytotoxin 2b of E. coli as highly specific antigens in ELISA performed in the serodiagnosis of infec- tions caused by VTEC in humans.

Materials and methods: The study population, used for characterization of the humoral immune response to the recombinant proteins, consisted of37 patients suspected for VTEC infection, mainly with clinical manifestation of HUS. Additionally serum samples from 78 clinically healthy persons and 96 patients with different bacterial infections (control group) were tested. Recombinant proteins were expressing in E. coli BL21 (DE3) using the pET-30 Ek/LIC expression vector (Novagen). Purification was accomplished by immobilized metal (Ni(2+)) affinity column chromatography (His-trap).

Results: The antibodies against recombinant proteins were detected using the ELISA in about half of the tested patients suspected in clinical investigation for VTEC infection. Most of the antibodies belong to the IgG and IgA class of immunoglobulins. Statistical analysis of the results showed that the frequency of detecting antibodies among patients with HUS was significantly higher in relation to the clinically healthy persons. However, the percentage of positive results in the control group were also much higher than in healthy persons what may indicate for presence of non-specific reactions. The least non-specific response was detected by ELISA with the protein Tir as antigen.

Conclusions: The study showed that recombinant proteins Tir, intimin and verocytotoxin 2b of E. coli may be used as antigens in routine diagnosis of VTEC infections. The most specific antigen is a recombinant protein Tir.

Introduction: The aim of the study was a retrospective analysis of intestinal flora for the presence of multidrug-resistant strains, isolated from patients hospitalized in clinics Oncology Center from 01.01.2010 to 30.09.2015 r.

Methods: The multi-resistant strains were isolated from stool and rectal swabs. In order to increase the potential of multiple-resistant strains, the material was plated on the appropriate substrate. Determination of resistance mechanisms performed by general recommendations.

Results: Results of this study showed among isolated multiple-resistance strains a high proportion of Enterobacteriaceae strains producing β-lactamase mainly ESBL. Klebsiella pneumoniae consist of 31.9% of isolated strains, E. coli 28.74% and Enterococcus faecium VER -21.15%.

Conclusions: It is important to determine the microbiological status of hospitalized patients because colonized gastrointestinal tract multi-resistant strains may be one of the sources of serious infections.

Introductions: According to clinical observations primary neoplastic lesions in the head and neck are often complicated by infection. The incidence of postoperative complications of cancer lesions of the head and neck, oral cavity, upper airways or larynx is from 19% up to 47% cases but in extensive and long-term operations can reach 80%.

Methods: We retrospectively evaluated results of microbiological investigations of 312 clinical specimens collected from patients from Clinical Head and Neck Cancers of On- cology Center in Warsaw in the years 2008 -2012. All samples of clinical materials were plated on the suitable culture media and incubated according to the recommendations.

Results: The study has shown that from all collected clinical specimens 491 strains have been isolated. Nearly half of the cultured bacteria were Gram-positive cocci (48.7%), mainly methicillin-sensitive Staphylococcus aureus. Most Gram-negative bacteria isolated from neoplastic lesions in the head and neck belong to Enterobacteriaceae (21%), 23,4% of cultured bacteria grown anaerobically and they were mainly Gram-negative rods.

Conclusions: The presence of diverse bacterial flora colonizing the mucous membranes of the mouth and throat may be the reason of difficulties in interpretation of microbiological investigation. It must be remembered that knowledge of colonizing and pathogenic flora of the area of head and neck enables appropriate preoperative prevention and empiric therapy.

the analyzed peritoneal fluid and bile specimens were comparable. Multiple bacterial species were significantly more common in bile isolates than in peritoneal fluid isolates. A total of 61,7% of aerobic Gram-negative bacillus isolates obtained from peritoneal fluid and bile produced ESBL. The proportions of vancomycin-resistant enterococci (VRE) and enterococci exhibiting high-level aminoglycoside resistance (HLAR) were 32,6% and 43,5%, respectively. Ertapenem-resistant K. pneumoniae was detected in 22,2% of peritoneal fluid cultures and 71,4% of biliary cultures. Methicillin resistance was detected in 85,7% of staphylococcal isolates. The proportion of anaerobes detected in peritoneal fluids was relatively high at approximately 17% and included predominantly Gram- negative species. All Gram-negative anaerobes showed resistance to benzylpenicillin. Conclusions: Etiologies and susceptibility pattern of IAls must be monitored on a ward, hospital, regional, and world-wide scale and the findings implemented into epidemiologic surveillance programs and proposed treatment protocols.

Introduction: Clostridium perfringens strains that produce enterotoxin often cause outbreaks of food poisoning or epidemic, therefore is essential to develop and adapt methods useful in epidemiological investigations. The aim of the study was to assess the genetic diversity of C. perfringens strains isolated from patients with symptoms of food poisoning and food samples.

Methods: In this study it has been determined the genetic diversity of the pilot group of epidemiologically non- related 39 C. perfringens strains using RAPD (Random Amplified Polymorphic DNA), rep-PCR (Repetitive DNA Element Based PCR) and PFGE (Pulsed Field Gel Electrophoresis) methods.

Results and conclusions: It has been obtained comparable results of differentiation of C. perfringens strains isolated from patients with symptoms of food poisoning and food samples using RAOD and rep-PCR methods. It has been also confirmed a wide genetic diversity among C. perfringens strains isolated from different sources epidemiologically non-related using PFGE.

Introduction: In the recent years there has been an increase in the incidence of infectious diseases that may be caused by the lack of guardians' consent to carry out immunization among children. In Poland in 2012 there was an increase in the incidence of pertussis 180% higher than in 2011. Occurring symptoms of pertussis can occur with varying intensity depending on the patient's age, his general health and the number of taken doses of vaccination against pertussis. The aim of the study was the analysis of reported in the years 2004 - 2013 confirmed cases of pertussis in the district of Bielsko and the city Bielsko- Biala, regarding to the prevalence of disease, the age of patients and the current course of compulsory vaccination against pertussis.

Methods: The analysis comprised 110 people with confirmed infection with Bordetella pertussis. The analysis included data from the years 2004-2013.

Results: On the basis of the analysis of the results the highest number of the patients with pertussis was reported in 2012. Women suffered more often than men. The median of the age of patients was 14 years. Pertussis was diagnosed in 7 infants, of which 5 have not yet received vaccination against pertussis. After 17 years of age incidence of pertussis occurred sporadically and involved 1-2 cases in each age group. 15,5% of patients had mandatory vaccination, and 52,7% of patients who suffered from pertussis received all doses of the primary vaccination, compulsory to the second year of life.

Conclusions: Between 2004-2013 in the district of Bielsko and the city Bielsko- Biala recorded 110 cases of pertussis, with the highest incidence in 2012. The largest group of patients were adolescents aged 14-16 years who have had vaccinations against pertussis, in force until the second year of life.

Introduction: A fungal infection of vagina is one of the most common gynecological problems of women of childbearing age. There has been a gradual increase in yeast- like fungi which are problematic to identify. There is thus a need for searching new identification methods, which would allow for fast recognition and selection of the proper course of treatment. The aim of study was to evaluate the usefulness of MALDI-TOF mass spectrometry technique in the diagnosis of pathogenic yeast-like fungi isolated from the genital tract of women.

Methods: There were identified 484 yeast-like fungi strains isolated from the genital tract of patients treated at Centrum Badah Mikrobiologicznych i Autoszczepionek im. Jana Bobra in Cracow (CBMiA) in the age group 20 - 50. The material was analyzed using streak plating on the culture medium: Sabouraud agar (CBMiA) and chromogenic media chromID Candida (bioMdrieux). The strains were identified using MALDI-TOF mass spectrometer (MALDI Biotyper MicroflexTM Series, Bruker Daltonics, Germany) operating as IVD (In Vitro Diagnostic). The procedure of proteins extraction with the use of ethanol and formic acid was used.

Results: All 484 yeast-like fungi strains isolated from the genital tract of women were correctly identified to the species level.

Conclusions: MALDI-TOF mass spectrometry method may be successfully used for routine identification of yeast-like fungi and in particular the strains of non-albicans Candida, which have a lower susceptibility to antifungal agents.

Introduction: Immunodeficient patients, e.g. transplant recipients, patients treated with corticosteroids, people with AIDS and individuals undergoing prolonged antibiotic therapy are at high risk of invasive fungal infections, especially invasive aspergillosis. Basic method for detection of organ/systemic fungal infection is serological monitoring in body fluids, first of all in serum, bu also in broncho-alveolar lavages (BALF). Proven invasive fungal infection should be diagnosed by culture of the pathogen or histopathological examination of infected tissues, however the detection of soluble fungal antigens in body fluids gives enough information for diagnosis of probable fungal infection, according to European Organization for Research and Treatment of Cancer recommendations, what allows introduction of antifungal therapy. Aim of the study was to asses the frequency of detection of circulation soluble fungal antigens with use of immunoenzymatic techniques in patients hospitalized between 2010 and 2015 in Independent Public Central Clinical Hospital (IPCCH) in Warsaw.

Methods: In IPCCH, between 2010 and 2015, 6475 serum samples, taken from 2096 patients, was tested for Candida spp. mannan antigen, and 7745 sera from 2243 patients were tested for Candida spp. mannan antigen, and 7745 sera from 2243 patients were tested for galactomannan antigen of Aspergillus spp, as well as 64 samples of BALF. Material was collected mainly from haematopoietic stem cell transplant recipients, hospitalized in Haematology and Oncology Clinics, during their routine pos-transplant monitoring. Testing was performed with use of quantitative (Candida antigen) or semiquantitative (Aspergillus antigen) immunoenzymatic methods (BioRad-Platelia), according to respective protocols.

Results: During examined period, increase in number of examinations was observed, starting from 1311 tests performed in 2010, up to 3052 examination in 2015. In 2015 testing for Aspergillus antigen in BALF samples was also introduced, resulting in 64 samples tested. Candida spp. antigen was detected in 171 samples (2,7% of all tested samples), and Aspergillus galactomannan was detected in 645 serum samples (8,4%) and 8 BALF samples (12,5%). Majority of examinations was performed for patients hospitalized in Haematology and Oncology Clinics (72,7%), Blood Vessel Surgery and Transplantology Clinics (3,8%), as well as in patients under care of post-transplantation (8,3%) and haematology (4,2%) out-patients clinics.

Conclusions: (i) In the 2015-2015 visible increase in number of fungal antigens examinations was observed, (ii) significant number of examinations was performed in onco-haematological patients (88,7%), what also indicates main risk group, (iii) 8,3% of fungal antigen testing was performed in solid organ transplant recipients, the second risk group for invasive fungal infection.

Despite the enormous development of vaccinology in recent decades, vaccinations of preg- nant women are still controversy. According to data from the literature, most of them are not only effective but also safe. The paper discusses the issues of vaccination among preg- nant women, with special accent on the recommendations of the most important Institu- tions of Public Health for this group of women.