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PL
Oceniono odczynem western-immunoblotting częstość występowania przeciwciał dla poszczególnych wydzielniczych białek Yop w próbkach surowicy osób z różnymi postaciami jersiniozy. Najczęściej w próbkach surowicy osób z objawami bólów brzucha, zapalenia stawów i rumienią guzowatego wykrywano przeciwciała dla białka YopD o masie 36 kDa. Przeciwciała klasy IgG dla białek YopD, YopM i YopH występowały w podobnym odsetku u osób z objawami bólów brzucha i zapaleniem stawów, przeciwciała klasy IgA dla tych białek dominowały u osób z zapaleniem stawów natomiast przeciwciała klasy IgM u osób z objawami bólami brzucha.
EN
The results obtained with the use of the western-blotting showed that antibodies for released proteins YopD (33-36 kDa) were the most frequently detected antibodies in serum samples from patients suspected for yersiniosis. Reactions between serum samples studied and the YopD proteinwere very intense, suggesting that protein is the strongest immunogen among the utilised, released proteins Yop of Yersinia. Antibodies IgM were more often diagnosed in patients with abdominal pain in the contrary to antibodies IgA which were characteristic to patients with reactive arthritis. Detailed analysis of the results of western-blotting on serum samples obtained several times from individuals with yersinosis during the course of infection in this investigation have showed also that antibodies of the IgA class hold longer in serum of individuals with arthritis compared with individuals with yersinosis not complicated by arthritis. In joint-fluid samples obtained from patients with arthritis antibodies for particular released proteins Yop were detected in the same class of immunoglobulins like in serum samples obtained from those individuals.
PL
Analiza wyników badań próbek surowicy uzyskanych od osób chorych podejrzanych o jersiniozę wykazała, że humoralna odpowiedź zależna jest od objawów klinicznych, wieku badanych osób, płci i okresu choroby. Najczęściej, bez względu na dominujące objawy kliniczne, wykrywano przeciwciała dla antygenu 03 Y. enterocolitica w badanych klasach immunoglobulin A, G i M, najrzadziej zaś dla antygenu 08 pałeczek tego gatunku.
EN
The antibodies against the somatic antigens of Y. enterocolitica 03, 08, 09, 05,27,Y. pseudotuberculosis I, and released proteins Yop were detected using the ELISA in 1634 serum samples and 84 synovial fluids collected from 1290 persons suspected for yersiniosis, as well as 200 serum samples from healthy individuals (blood donors). The presence of antibody in diagnostically significant titres for somatic antigens of Yersinia were detected by ELISA in 20.5% and 50.6%, antibodies for released proteins Yop in 11.5% and 28.4% respectively of blood donors and patients suspected for yersiniosis. The antibody against the 03 antigen of Y. enterocolitica was the most frequently detected antibody while the most infrequent was the antibody for the antigen from the 08 serologic group. The results of the study showed that the humoral response picture to Yersinia antigens in the course of yersiniosis in humans is dependent on the age and sex of the patient, duration of the infection, and clinical manifestations. Most frequently the elevated antibody levels were detected among patients with erythema nodosum and patients with gastrointestinal symptoms. The frequency of occurrence of antibodies for most antigens of Yersinia, together with age increased reaching its peak, on the average, among individuals aged 21-40 years. Analysis of individual cases showed that by the end of the first week of infection, elevated levels of antibodies for somatic antigens of Yersinia are evident. On the other hand, antibodies for released proteins Yop as a matter of rule appear in the second week from the onset of clinical symptoms. Within this early phase of infection immunoglobulins of the A and M classes dominate reaching their highest level in the second to third week of the infection. In majority of the individuals studied antibodies of the IgG class reached their highest level much later in relation to those of the IgA and IgM classes. Significant differences were found in IgA antibody detection among individuals with clinical manifestations of stomachaches and arthritis. Nevertheless, among individuals with clinical symptoms of stomachaches, these immunoglobulins as a matter of principle disappear with a period of 2-3 months from the onset of clinical symptoms. In individuals with arthritis however the aforementioned immunoglobulins maintained at considerable levels even after a year. In joint-fluid samples obtained from patients with arthritis antibodies for Yersinia antigens were detected in similar levels just as obtained simultaneously serum from those individuals.
EN
Shiga toxin-producing Escherichia coli (STEC) strains also called verotoxin-producing E. coli (VTEC) represent one of the most important groups of food-borne pathogens that can cause several human diseases such as hemorrhagic colitis (HC) and hemolytic – uremic syndrome (HUS) worldwide. The ability of STEC strains to cause disease is associated with the presence of wide range of identified and putativevirulence factors including those encoding Shiga toxin. In this study, we examined the distribution of various virulence determinants among STEC strains isolated in Poland from different sources. A total of 71 Shiga toxin-producing E. coli strains isolated from human, cattle and food over the years 1996–2010 were characterized by microarray and PCR detection of virulence genes. As stx1a subtype was present in all of the tested Shiga toxin 1 producing E. coli strains, a greater diversity of subtypes was found in the gene stx2, which occurred in five subtypes: stx2a, stx2b, stx2c, stx2d, stx2g. Among STEC O157 strains we observed conserved core set of 14 virulence factors, stable in bacteria genome at long intervals of time. There was one cattle STEC isolate which possessed verotoxin gene as well as sta1 gene encoded heat-stable enterotoxin STIa characteristic for enterotoxigenic E. coli. To the best of our knowledge, this is the first comprehensive analysis of virulence gene profiles identified in STEC strains isolated from human, cattle and food in Poland. The results obtained using microarrays technology confirmed high effectiveness of this method in determining STEC virulotypes which provides data suitable for molecular risk assessment of the potential virulence of this bacteria.
PL
Przeprowadzono elektroforetyczne i serologiczne badania porównawcze składu białek 6 szczepów M. pneumoniae i 1 szczepu M. genitalium. Stwierdzono pełne podobieństwo budowy antygenowej badanych szczepów M. pneumoniae oraz bliskie podobieństwo antygenowe M. genitalium do M. pneumoniae. Najbardziej swoistymi spośród antygenów M. pneumoniae okazały się białka o masie cząsteczkowej 170 (białko P1) i 89 kDa, uważane za najważniejsze adhezyny komórki mykoplazmowej.
EN
The Mycoplasma pneumoniae FH strain routinely used in our laboratory for over 25 years as antigen in serological tests, 2 reference M. pneumoniae strains from ATCC (29342 and M129) and 3 isolates of M. pneumoniae obtained in 1995 from pneumonia patients were compared by SDS-PAGE, complement fixation test (CFT) and by Western-immunoblotting against human and rabbit serum samples with high level of mycoplasmal antibodies. On SDS-PAGE all M. pneumoniae strains showed the same number of 23 polypeptides on the gel with identical molecular weights. The same strains on immunoblotting against human and rabbit serum samples showed six bands: 170, 89, 75, 55, 38 and 33 kDa with the strongest antibody staining in 170-(P1 protein) and 89-kDa bands. Because of its known antigenic relationships Mycoplasma genitalium was used for comparison. The pattern of M genitalium proteins on SDS-PAGE was similar to pattern of M. pneumoniae but distinguishable. On immunoblotting six proteins of M. genitalium (135, 127, 110, 95, 75 and 45 kDa) reacted with human and rabbits immunoglobulins for M. pneumoniae antigens. Furthermore in complement fixation test both antigens, prepared from M. pneumoniae and M. genitalium, reacted as well with human and rabbit immunoglobulins for M. pneumoniae and with rabbit immunoglobulins for M. genitalium. These cross-reactions observed in serological techniques could give false positive results in routine diagnosis of M. pneumoniae infections. In such situations showing on immunoblott of presence in tested serum sample of antibodies to 170 - and 89 kDa proteins could confirm M. pneumoniae infection.
PL
Zbadano 173 próbki materiału klinicznego techniką PCR, przy użyciu komercyjnego testu Mycoplasma IST2 oraz klasyczną metodą hodowli w kierunku obecności U. urealyticum i M. hominis. Stwierdzono wysoką zgodność wyników uzyskanych we wszystkich 3 metodach przy poszukiwaniu U. urealyticum. W przypadku badania próbek materiału klinicznego w kierunku M. hominis wyniki dodatnie uzyskano wyłącznie przy zastosowaniu testu Mycoplasma IST2, co sugeruje, że mogą to być wyniki fałszywie dodatnie.
EN
The polymerase chain reaction (PCR) technique and commercial Mycoplasma IST 2 test were compared with culture for the detection of U. urealyticum and M. hominis in 173 clinical samples obtained from patients without clinical symptoms from genito-urinary tract. The presence of U. urealyticum was diagnosed by culture in 24 samples, by PCR in 33 samples and by Mycoplasma IST 2 test in 39 samples. The presence of M. hominis was diagnosed in 26 samples only by Mycoplasma iST 2 test - culture and PCR were negative. The study showed the excellent sensitivity (100%) and good specificity (appropriately 94.0% and 90.0%) for U. urealyticum in PCR and Mycoplasma 1ST 2 test. The discrepancy of results obtained in Mycoplasma IST 2 test and culture as well as in PCR may suggest the over sensitivity of the copimercial test for detection of M. hominis.
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