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Tytuł artykułu

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

Autorzy

Warianty tytułu

Języki publikacji

PL

Abstrakty

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.

Wydawca

-

Rocznik

Tom

52

Numer

1

Opis fizyczny

s.35-49,fot.,tab.,bibliogr.

Twórcy

  • Panstwowy Zaklad Higieny, 00-791 Warszawa, ul.Chocimska 24

Bibliografia

  • 1. Bottone EJ. Yersinia enterocolitica: the charisma continues. Clinic Microbiol Rev 1997; 10: 257-76.
  • 2. China B, N'Guyen BT, De Bruyere M, Cornells GR. Role of YadA in resistance of Yersinia enterocolitica to phagocytosis by human polymorphonuclear leukocytes. Infect Immun 1994; 62: 1275-81.
  • 3. Comelis GR, Sluiters Ch, Lambert De Rouvrait C, Michieh T. Homology between virF, the transcriptional activator of the Yersinia virulence regulon, and araC, the Escherichia coli arabinose operon regulator. J Bacteriol 1989; 171: 254-62.
  • 4. Feng P, Keasler SP, Hill WE. Direct identification of Yersinia enterocoiitica in blood by polymerase chain reaction amplification. Transfusion 1992; 32: 850-4.
  • 5. Gierczyński R, Ocena przydatności wybranych markerów wirulencji do identyfikowania chorobotwórczych szczepów pałeczek Yersinia enterocolitica. I. Fenotypowe markery związane z plazmidem pYV. Med Dośw Mikrobiol 2000; 52: 25-34.
  • 6. Gierczynski R, Jagielski M, Szych J, Kałużewski S. Występowanie plazmidów wirulencji w szczepach pałeczek Yersinia pochodzących z różnych źródeł. Med Dośw Mikrobiol 1996; 48: 141-50.
  • 7. Górka A, Brzostek K, Hrebenda J. Białka Yop rodzaju Yersinia - mechanizm sekrecji i translokacji. Post Mikrobiol 1999; 38: 119-41.
  • 8. Heesemann J i inni. Plasmids of human Strains of Yersinia enterocolitica: molecular relatedness and possible importance for pathogenesis J Infect Dis 1983; 147: 107-15.
  • 9. Kapperud G, Namork E, Skumik M, Nesbakken T. Plasmid-mediated surface fibrillae of Yersinia pseudotuberculosis and Yersinia enterocolitica: relationship to the outer membrane protein YOP1 and possible importance for pathogenesis. Infect Immun 1987; 55: 2247-54.
  • 10. Kapperud G, Nesbakken T. Restiction endonuclease analysis of 40 to 50 Mdalton plasmids from Y. enterocolitica strains of worldwide origin. Cont Microbiol Immunol 1987; 9: 317-23.
  • 11. Kapperud G, Vardund T, Skjerve E i inni. Detection of pathogenic Yersinia enterocoiitica in foods and water by immunomagnetic separation, nested polymerase chain reactions, and colorimetric detection of amplified DNA. Appl Environ Microbiol 1993; 59: 2938-44.
  • 12. Kur J. Podstawy inżynierii genetycznej. Wydawnictwa Politechniki Gdańskiej, Gdańsk, 1994.
  • 13. Lian CJ, Hwang WS, Pai CH. Ptasmid-mediated resistance to phagocytosis in Yersinia enterocolitica. Infect Immun 1987; 55: 1176-83
  • 14. Mullis K Faloona F, Scharf S i inni. Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. Cold Spring Harbor Symp. Quant Biol 1986; 51: 263-73.
  • 15. Pilz D, Vocke T, Heesemann J, Bra V. Mechanism of YadA-mediated serum resistance of Yersinia enterocoiitica serotype 03, Infect. Immun., 1992; 60: 189-95.
  • 16. SambrookJ, Fritsch E, Maniatis T. Molecular cloning. A laboratory manual, red. Cold Spring Harbor Lab. Press. 1989.
  • 17. Skumik M, Wolf-Watz H. Analysis of the yopA gene encoding the Yop1 virulence determinants of Yersinia spp. Mol Mirobiol 1989; 3; 517-29.
  • 18. Straky SC, Piano GV, Skrzypek E, Bliska JB. Yops of Yersinia spp. Pathogenic for humans. Infect Immun 1993; 61: 3105-10.
  • 19. Wiśniewski J, Bielecki J. Mechanizmy wirulencji bakterii z rodzaju Yersinia. Post Mikrobiol, 1996; 35: 213-241.
  • 20. Wren BW, Tabasqchali S. Detection of pathogenic Yersinia enterocolitica by the polymerase chain reactin. Lancet, 1990; 336: 693.

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

bwmeta1.element.agro-article-84db8c1c-2956-427e-99d9-f2152c93b0bc
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