Wyniki wyszukiwania - Biblioteka Nauki

1

Ocena przydatnosci wybranych markerow wirulencji do identyfikowania chorobotworczych szczepow paleczek Yersinia enterocolitica. Cz.II. Genotypowe markery zwiazane z plazmidem pYV

100%

Gierczynski R.

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tom 52

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nr 1

35-49

PL

Wykrycie w genomie pałeczek Y. enterocolitica genów virF i yadA związanych z plazmidem wirulencji (pYV) pozwala na identyfikację chorobotwórczych szczepów tego drobnoustroju izolowanych z materiału klinicznego od ludzi.

EN

Pathogenic strains of Yersinia enterocolitica bear virulence associated plasmid pYV. Unfortunately plasmid pYV is easily lost by these bacteria incubated at elevated temperatures (37°C) or long stored at room temperatures. This sometimes makes difficult the detection of the virulence plasmid, especially by its isolation or biochemical tests. On the other hand, observations done by some authors suggest that polymerase chain reaction (PCR) could be useful for demonstration of the pYV plasmid of Yersinia strains. Accordingly to this observation the aim of the presented study was to check the usefulness of plasmid-localised genes virF and yadA, detected by PCR, for the identification of the virulent strains of Y. enterocolitica. In the presented study one hundred and fifty two clinical strains of Y enterocolitica belonging to serogroup O3 were investigated by the PCR for the presence of genes virF and yadA. Bacterial strains were first tested for the presence of pYV plasmid. In addition the phenotypic features: calcium dependence, Congo red binding and autoagglutination were determined. In this way the virulence plasmid was found in 130 of 152 examined strains. For PCR studies also forty plasmid-cured strains of K enterocolitica and 32 non - Y. enterocolitica, Enterobacteriaceae strains were included. The obtained results show that the tested genes were present only in Yersinia strains possessing the pYV plasmid and no one non-specific PCR product was observed. The detection level of these genes in nested PCR permits to detect pathogenic Y. enterocolitica in suspension composed of 1 x 10(9) CFU/ml of pYV+ bacilli and 3 x 10(9) CFU/ml plasmid-cured, isogenic bacteria. In the study it was shown that genes virF and yadA were useful virulence markers, which could be helpful in clinical studies for the detection of the virulence plasmid in Y. enterocolitica strains long stored or incubated at elevated temperatures.

2

Diagnostyka i molekularna epidemiologia Bacillus anthracis

100%

Gierczynski R.

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nr 3

165-172

3

Filogeneza, chorobotwórczość i zróżnicowanie pałeczek Yersinia enterocolitica

100%

Gierczynski R.

Medycyna Doświadczalna i Mikrobiologia

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2012

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tom 64

|

nr 2

4

Heterogeneity of galF and gnd of the cps region for capsule synthesis in clinical isolates of Klebsiella pneumoniae

100%

Gierczynski R.

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tom 56

|

nr 2

EN

The capsular polysaccharide (CPS) plays important role in Klebsiella spp pathogenesis. Capsular types Kl and K.2 of Klebsiella pneumoniae are considered most virulent for humans. The capsule biosynthesis region flanking genes galF and gnd from clinical isolates and reference strains of K. pneumoniae were screened for polymorphism. Nucleotide sequence analysis of galF and gnd revealed a high heterogeneity. However, deduced amino acid sequences demonstrated that the majority of mutations were silent implying GalF and Gnd are strongly conserved. This may suggest importance of these loci in the CPS biosynthesis and may argue for their potential usefulness in Klebsiella genotyping.

5

Ocena przydatnosci nowych enzymow restrykcyjnych SfaAI oraz SmiI do roznicowania izolatow Yersinia enterocolitica 4-O3 i 1B-O8 metoda elektroforezy w zmiennym polu elektrycznym [PFGE]

63%

Zacharczuk K. , Gierczynski R.

Medycyna Doświadczalna i Mikrobiologia

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2009

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tom 61

|

nr 2

133-142

6

Elektroforetyczna i immunologiczna analiza natywnych białek wydzielniczych wytwarzanych in vitro w warunkach indukcji Ysa (Yersinia secretion apparatus) przez izolaty Yersinia enterocolitica 1B/O8 wyosobnione od ludzi w Polsce

51%

Rokosz-Chudziak N. , Rastawicki W. , Zacharczuk K. , Gierczynski R.

Medycyna Doświadczalna i Mikrobiologia

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2013

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tom 65

|

nr 4

7

Porownanie lancuchowej reakcji polimerazy [PCR] oraz komercyjnego testu Mycoplasma IST2 z klasyczna hodowla bakteryjna w diagnostyce zakazen wywolywanych przez Ureaplasma urealyticum i Mycoplasma hominis

51%

Rastawicki W. , Kalota H. , Jagielski M. , Gierczynski R.

Medycyna Doświadczalna i Mikrobiologia

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2004

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tom 56

|

nr 1

99-108

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.

8

Extended multiple-locus variable-number tandem-repeat analysis of Bacillus anthracis strains isolated in Poland

51%

Gierczynski R. , Jakubczak A. , Jagielski M.

Polish Journal of Microbiology

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2009

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tom 58

|

nr 1

3-7

EN

Twenty-one variable-number tandem-repeat (VNTR) marker loci were used for extended multiple locus VNTR analysis (MLVA) of 14 laboratory strains of Bacillus anthracis isolated in Poland and vaccine strain Sterne 34F2A. The extended MLVA (MLVA-21) distinguished six genotypes clustered in three main branches. Monomorphic branch 1 consisted of the vaccine strain and six isolates from distinct samples of a cow died from anthrax. This group also encompassed three haemolytic isolates of B. anthracis. Branches 2 and 3 were heterogeneous and consisted of five and three isolates of the phylogenetic lineages B2 and A1, respectively. MLVA-21 supported thesis on the anthrax agent heterogeneity in Poland. This study brought an additional evidence that haemolytic B. anthracis strains isolated in Poland are closely related to the vaccine strain Steme 34F2 and may together constitute the same sensu stricto strain. No epidemiological link could be however traced between both the vaccine and the haemolytic strains.

9

Wystepowanie plazmidow wirulencji w szczepach paleczek Yersinia pochodzacych z roznych zrodel

51%

Gierczynski R. , Jagielski M. , Szych J. , Kaluzewski S.

Medycyna Doświadczalna i Mikrobiologia

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1996

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tom 48

|

nr 3-4

141-150

10

Zastosowanie metody do wykrywania wybranych genow Mycoplasma pneumoniae

51%

Rastawicki W. , Jagielski M. , Gierczynski R.

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tom 52

|

nr 1

67-74

PL

Wystandaryzowano technikę łańcuchowej reakcji polimerazy (PCR) do wykrywania genów M. pneumoniae kodujących białko PI, pod jednostkę 16S rRNA i czynnik ekongacyjny Tu. Stwierdzono, że PCR jest techniką umożliwiającą wykrycie poszukiwanych genów nawet wtedy, gdy w 1 ml hodowli znajduje się od 10(2) do 10(4) komórek M. pneumoniae.

EN

The aim of this study was standardization of PCR for the detection of gene encoding the PI protein, 16S rRNA and elongation factor Tu of M. pneumoniae. A total of 13 strains of M. pneumoniae, 28 strains of other mycoplasmas and 14 strains of different bacteria causing respiratory tract infections were tested. In all of tested M. pneumoniae strains the presence of the sought genes was confirmed. The specificity of DNA was confirmed by the restriction endonuclease analysis with enzymes Hind III, Alu I and Hha I. With none of primers specific for the M. pneumoniae genes amplification of DNA from other bacteria was noted. The PCR method with the selected primers allowed to detect from 10(2) to 10(4) cfu M. pneumoniae/ml suspended in broth. The obtained results indicate that the PCR method can be used for detection of M. pneumoniae genes. A very good sensitivity and specificity predestine PCR as a potential quick diagnostic method for identification of M. pneumoniae in clinical specimens.

11

Wykorzystanie wydzielniczych bialek Yop paleczek Yersinia w serologicznej diagnostyce jersiniozy. II. Zastosowanie metody rekombinacji genetycznej do uzyskania bialka YopD

51%

Rastawicki W. , Gierczynski R. , Jagielski M.

Medycyna Doświadczalna i Mikrobiologia

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2004

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tom 56

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nr 1

41-47

PL

Metodami rekombinacji genetycznej uzyskano preparat wydzielniczego białka YopD, który użyto jako antygenu w odczynie ELISA do serologicznej diagnostyki jersiniozy. Wyniki oznaczeń poziomu surowiczych przeciwciał dla tak uzyskanego antygenu pałeczek Yersinia porównano z wynikami oznaczeń uzyskanymi przy użyciu testu ELISA, w którym jako antygenu użyto białek wydzielniczych uzyskanych z supernatantu hodowli Y enterocolitica oraz przy użyciu komercyjnego testu ELISA z rekombinowanym antygenem białkowym. Na podstawie wyników oznaczeń wykazano pełną przydatność uzyskanego we własnym zakresie metodą rekombinacji genetycznej preparatu wy- dzielniczego białka YopD do diagnostyki jersiniozy.

EN

The aim of this study was to evaluate the usefulness of gene recombination technique using the pET-30 Ek/LIC expression vector for production a 36 kDa released protein called YopD and evaluate of this purified protein as antigen in serodiagnosis of yersiniosis. Protein YopD of Y enterocolitica was expressing in Escherichia coli BL21 (DE3) using the pET-30 Ek/LIC expression vector. Purification of the expressed enzyme from suspensions of E. coli cells treated with Bug BusterTM Protein/Extraction Reagent was accomplished by immobilised metal (№2+) affinity column chromatography (His-trap). The IgM, IgG and I gA class antibodies to YopD were measured in 100 serum samples collected from patients suspected for yersiniosis and 100 blood donors. The obtained results were compared to the results of ELISA with released proteins isolated from the culture of Y enterocolitica supernatant under calcium deficient conditions and commercial ELISA with recombinant released proteins. A very high (94.0-100.0%) specificity and good sensitivity (55,2 - 80.4%) were displayed by the ELISA with YopD in relation to other two ELISA. The results of our study showed that recombinant YopD protein purified by chromatography of bio-affinity may be used in serodiagnosis of yersiniosis as a high specific antigen free of Yersinia lipopolysaccharides.

12

Zastosowanie metody inżynierii genetycznej do uzyskania białka P1 Mycoplasma pneumoniae oraz ocena jego przydatności w serodiagnostyce mykoplazmozy u ludzi

51%

Rastawicki W. , Rokosz N. , Gierczynski R. , Jagielski M.

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nr 3

13

Wystepowanie i charakterystyka szczepow opornych na antybiotyki oksy-imino-betalaktamowe wsrod szczepow Salmonella enterica subsp. enterica izolowanych w Polsce

51%

Szych J. , Gierczynski R. , Wardak S. , Cieslik A.

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nr 2

115-130

PL

Scharakteryzowano 68 szczepów pałeczek Salmonella opornych na antybiotyki oksy-imino-betalaktamowe należących do 6 typów serologicznych, wybranych spośród 239 opornych na ampicylinę szczepów pałeczek Salmonella izolowanych w Polsce w latach 1999-2004 z próbek materialu klinicznego. Stwierdzono, że większość badanych szczepów wytwarzała p-Iaktamazę TEM-1 o pl=5,4 oraz p-laktamazę o rozszerzonym spektrum substratowym CTX-M-3 o pl=8,4. Wszystkie badane szczepy S. Thompson i jeden ze szczepów S. Typhimurium wytwarzał enzym SHV-5 o pl=8,2. Jest to pierwsze doniesienie o produkcji SHV-5 przez pałeczki Salmonella w Polsce.

EN

Extended - spectrum ß-lactamases (ESBLs) are enzymes manifesting considerable hydrolyzing activity on a wide variety of ß-lactam antibiotics including oxyiminocephalosporins and aztreonam. In the study reported here we investigated the types of ESBL produced by Salmonella enterica subsp. enterica strains isolated from clinical samples in the microbiological laboratories of sanitary-epidemiological units in Poland from 1999 to 2004. Among 239 ampicilline-resistant Salmonella enterica subsp. enterica strains isolated from clinical samples in the microbiological laboratories of sanitary-epidemiological units in Poland, 68 isolates of oximino-beta-lactams resistant of 6 serovars were found. There were 16 epidemiological unrelated strains (6 isolates of S.Enteritids, S isolates of S.Thompson, 3 isolates of S.Typhimurium, onefold isolate of S.Muenster and S. enterica 1,9,12:-:-) coming from different areas of country and 52 epidemiologically related isolates of S. Oranienburg, coming from a prolonged outbreak in an orphanage in Łódź. All the strains were identified as the ESBLs producers . The molecular analysis relevead that most of them expressed CTX-M-3 ESBL which is widely observed in Poland and additional enzyme TEM-1. All tested isolates of S.Thompson and one of three S. Typhimurium isolates were found to produce SHV-5 ESBL. This is the first report regarding the presence of SHV-5 in the genus Salmonella in Poland.

14

Wykorzystanie wydzielniczych bialek Yop paleczek Yersinia w serologicznej diagnostyce jersiniozy I.Antygen do odczynu immunoenzymatycznego

51%

Rastawicki W. , Jagielski M. , Gierczynski R.

Medycyna Doświadczalna i Mikrobiologia

|

2003

|

tom 55

|

nr 2

125-133

PL

Z hodowli pałeczek Y enterocolitica w podłożu pozbawionym jonów wapnia uzyskano preparat wydzielniczego białka Yop, który użyto jako antygen w odczynie ELISA do serologicznej diagnostyki jersiniozy. Wyniki oznaczeń poziomu surowiczych przeciwciał dla tak uzyskanego antygenu pałeczek Yersinia porównano z wynikami oznaczeń uzyskanymi przy użyciu komercyjnych testów firmy Mikrogen i Euroimmun. Wykazano pełną przydatność uzyskanego we własnym zakresie preparatu wydzielniczych białek Yop do serologicznej diagnostyki jersiniozy.

EN

The usefulness of the released proteins (RPs) prepared in our laboratory from the strain Y enterocolitica for the serological diagnosis of yersiniosis was estimated. Yersinia enterocolitica 0:8 was cultured under calcium deficient conditions and the virulence factors were isolated from the culture supernatant. The purified proteins were sulubilized using sodium dodecyl sulfate. The results of ELISA using released proteins obtained in our laboratory as the antigen were compared to the results of commercial ELISA-Mikrogen and ELISA-Euroimmun. One hundred serum samples obtained from patients suspected in clinical investigations for jersiniosis were tested. Comparison of the frequency of detecting in particular ELISA antibodies to Yersinia showed a high ( > 85%) agreement of the results in all of the immunoglobulin classes. The highest sensitivity (97.4%) and specificity (95.4%) was displayed by the ELISA with our antigen in relation to the ELISA-Mikrogen when determining antibodies of IgM class.

15

Wykorzystanie wydzielniczych bialek Yop paleczek Yersinia w serologicznej diagnostyce jersiniozy. III. Odczyn western-immunoblotting

51%

Rastawicki W. , Jagielski M. , Gierczynski R.

Medycyna Doświadczalna i Mikrobiologia

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2004

|

tom 56

|

nr 4

335-342

16

Ocena przydatnosci aglutynacyjnego testu z wyciagiem Mangifera indica do identyfikowania chorobotworczych szczepow Yersinia enterocolitica

51%

Kaluzewski S. , Gierczynski R. , Szych J. , Jagielski M.

Medycyna Doświadczalna i Mikrobiologia

|

1997

|

tom 49

|

nr 3-4

177-186

EN

The study was performed on 137 Y. enterocolitica strains belonging to various serological groups, including 75 O3 group strains isolated form human clinical material. The agglutination test on slides was carried out on this strains using Mangifera indica extract of own production. Agglutinating preparation obtained from the seeds of M. indica agglutinated Y. enterocolitica organisms possessing the pVY plasmid and CRMOX+ phenotype in dilutions to 1,56 µm/ml. In identification tests conducted paralelly agglutination solution was used in concentrations of 100 and 10 µm/ml. All clones of Y. enterocolitica from 03 group from cultures at 37°C and with CRMOX+ phenotype possessing the pVY plasmid were agglutinated by the extract. Agglutination failed to develop in the cultures of these clones incubated at 25°C. Yersinia clones not containing the pVY plasmid with CRMOX" phenotype were resistant to agglutination. The virulence plasmid was found in 44 out of 75 strains of Y. enterocolitica O3 and was identified by restriction analysis after plasmid DNA digestion with Eco RI enzyme. The obtained results agreed with those of Wauters et al. in 1995 and confirmed the opinion of these authors on the usefulness of the test with M. indica agglutinin for the identification of virulent Y. enterocolitica strains.

17

Ocena przydatnosci wybranych markerow wirulencji do identyfikowania chorobotworczych szczepow paleczek Yersinia enterocolitica. IV. Geny myfA i ureC

51%

Gierczynski R. , Jagielski M. , Rastawicki W.

Medycyna Doświadczalna i Mikrobiologia

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2002

|

tom 54

|

nr 4

347-355

PL

Badano występowanie genów związanych z wytwarzaniem ureazy (ureC) i fimbrii Myf (myfA) u pałeczek Yersinia. Stwierdzono, że gen myfA związany z wytwarzaniem fimbrii Myf może być przydatny w diagnostyce mikrobiologicznej do identyfikowania chorobotwórczych pałeczek Y. enterocolitica, które utraciły plazmid wirulencji pYV. Natomiast gen ureC uznany został za nieprzydatny do tego celu, ze względu na jego występowanie w genomie niechorobotwórczych pałeczek Y. enterocolitica należących do biotypu 1A oraz pałeczek J', intermedia i Y. kristensenii.

EN

We check by polymerase chain reaction (PCR) the presence of gene ureC and myfA, encoding subunits of urease and Myf fimbriae, among clinical and food-originated strains of Yersinia to determine their usefulness as molecular virulence markers of Y. enterocolitica. The examinations were done on 130 clinical strains of Y. enterocolitica O:3/4 isolated in Poland from humans. All strains were obtained from stool and possessed the virulence plasmid pYV. In addition 40 isogenic, plasmid-cured strains were tested. The 52 strains including Y. enterocolitica (biotype 1A, 4, 2 and 1B), Y. pseudotuberculosis, Y. intermedia, Y. frederiksenii, Y. kristensenii, E. coli, Citrobacter, Shigella and Salmonella were used as controls. The PCR assay resulted in detection of genes: ureC and myfA in genomic DNA of all 130 tested clinical strains of Y. enterocolitica pYV+, as well as in plasmid cured strains. Furthermore, ureC was found in all tested strains of Y. enterocolitica biotype A1 and in one strain of Y. intermedia and Y. kristensenii. In contrast to ureC, myfA was detected only in strains of Y. enterocolitica considered as pathogenic.Obtained results show, gene myfA seems to be the reliable virulence marker of Y. enterocolitica, whereas ureC is not recommended for identification of pathogenic strains of this species.

18

Molekularna charakterystyka hemolizujacych szczepow Bacillus anthracis izolowanych w Polsce

51%

Gierczynski R. , Zasada A. A. , Kaluzewski S. , Jagielski M.

Medycyna Doświadczalna i Mikrobiologia

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2006

|

tom 58

|

nr 4

339-346

PL

W prezentowanej pracy podjęto próbę zróżnicowania szczepionkowego szczepu Sterne 34F2 i atypowych izolatów B. anthracis wyosobnionych w Polsce drogą wykorzystania 10 loci VNTR, a także sprawdzenia czy takie czynniki jak: anthrolizyna O (gen alo), cereolizyna (gen clo), hemolityczna enterotoksyna HBL (gen hblA) i mutacja typu stop w genie plcR mogą być związane z hemolityczną aktywnością tych izolatów. Potwierdzono klonalne pokrewieństwo szczepów hemolizujących i nie wykazujących tej cechy. Mimo, że nie zidentyfikowano czynnika genetycznego warunkującego zdolność do hemolizy badanych szczepów, to wykazano, że badane szczepy posiadały geny alo, clo i mutację typu stop w genie plcR przy braku obecności genu hblA. Zebrane informacje umożliwiły pełniejszą charakterystykę hemolizujących szczepów B. anthracis izolowanych na terenie kraju.

EN

Bacillus anthracis is generally considered non-haemolytic, when cultured on the solid media. However, strains capable to lyse sheep erythrocytes have been reported. Anthrolysin O, an orthologue of cereolysin was proposed as a putative haemolysin of B. anthracis. AIM: to determine whether anthrolysin O, haemolytic enterotoxin HBL and the pleiotropic regulator PlcR that activates antrholysin O production are associated with a haemolytic activity of B. anthracis strains isolated in Poland. MATERIAL: in total 8 B. anthracis strains - the fully virulent BL1 and seven the pX02 lacking strains including: a vaccine strain Sterne 34F2 together with three haemolytic and three non-haemolytic strains isolated from different samples of the same animal died from anthrax in Poland. METHODS: The haemolytic activity was detected using Columbia agar plates supplemented with 5% of sheep blood. Anthrolysin O, cereolysin and gene hblA encoding the key subunit of the HBL were detected by PCR. In addition, the plcR gene fragment containing the B. anthracis specific non-sense mutation was analysed by the DNA sequencing. Ten marker loci based MLVA genotyping was performed to distinguish tested strains. RESULTS: The alo gene encoding anthrolysin O was detected in both the haemolytic and non-haemolytic strains while hblA was absent. The B. anthracis specific plcR non-sense mutation was detected in both the groups of tested strains, suggesting that the haemolysis in tested strains may rather be conferred by the PlcR-independent factors. Moreover, haemolytic and non-haemolytic strains were indistinguishable by the MLVA. Obtained results may argue the haemolytic and non-haemolytic strains are isogenic and most probably a single mutational event is responsible for the haemolytic phenotype induction.

19

Wykorzystanie odczynu ELISA w serologicznej diagnostyce tularemii

51%

Rastawicki W. , Jagielski M. , Gierczynski R.

|

nr 4

425-429

PL

Oceniono przydatność w serologicznej diagnostyce tularemii odczynu ELISA z uzyskanym we własnym zakresie antygenem F. tularensis. Stwierdzono, że odczynem ELISA możliwe było wykrycie swoistych przeciwciał u osób chorych w mianie kilkakrotnie wyższym niż rutynowo wykonywanym odczynem aglutynacji probówkowej. Wyniki przeprowadzonych badań wskazują na pełną przydatność opracowanego odczynu ELISA w serodiagnostyce tularemii.

EN

The usefulness of the ELISA using as antigen prepared in our laboratory supernatant obtained after centrifugation of sonicated F. tularensis cell suspension was compared with the tube agglutination test with commercial available antigen. Paired serum specimens obtained from 6 patients with ulceroglandular syndrome of tularemia were tested in both tests. The cut-off limit of serum antibodies was set at mean antibody titre determined in the sera of 115 blood donors exceeded by three standard deviations. Antibodies to F. tularensis in diagnostically significant titre were detected in all 12 serum samples by both tests. However the titres obtained in ELISA were several times higher than in tube agglutination test. In the second serum sample the level of IgA and IgM was lower but the level of IgG higher than in the first sample. We could not observe any difference in the level of antibodies between paired serum specimens in tube agglutination test.

20

Ocena przydatności diagnostycznego systemu komercyjnego ID 32 E do identyfikacji wybranych pałeczek z rodziny Enterobacteriaceae i innych gram-ujemnych pałeczek fermentujących glukozę

51%

Szych J. , Kaluzewski S. , Cieslik A. , Gierczynski R.

Medycyna Doświadczalna i Mikrobiologia

|

2001

|

tom 53

|

nr 4