Identyfikacja i charakterystyka shigatoksycznych szczepow Escherichia coli wystepujacych w surowej wolowinie przy uzyciu sondy molekularnej oraz multiplex PCR

• Autorzy: Weiner M

Warianty tytułu

EN

Identification and characterization of Shiga toxin-producing Escherichia coli strains originating from raw beef by the use of a molecular probe and multiplex PCR

• Języki publikacji: PL

Abstrakty

EN

Shiga toxin-producing E. coli (STEC) are a serious cause of human diseases. The infections are mainly associated with strains belonging to serogroups O157, O26, O103, O111 and O113, while the main source of infection is contaminated food of animal origin (especially beef). In this study the molecular identification methods for STEC were used which enabled detection of STEC in raw beef using multiplex PCR (mPCR) assays, isolation of individual bacterial colonies with digoxigenin (DIG)-labeled DNA probe and characterization of the virulence markers by the use of mPCR. The molecular method was used to examine 272 raw beef samples obtained from slaughterhouses. After the mPCR-1 analysis 16 stx-positive samples (5.88%) were detected. The STEC isolates were then tested using the mPCR and PCR assays. Eight of them belonged to O157:H7 serotype by the presence of the rfbO157 and fliCн₇ genes. The stx₁ marker was observed in all E. coli O157:H7 and in four of the non-O157:H7 isolates tested. Moreover, the stx₂ gene was detected in all E. coli O157:H7 and in seven of the non-O157:H7, three of them also possessed the stx₁ marker. Eight of the rfbO157-negative STEC were examined with the mPCR-3 assay to detect the rfbO111, rfbO113 and O26wzx markers, three of them produced the characteristic amplicon for the O26 group. These isolates also carried the stx₁ (one strain) or stx₂ genes (two isolates). Among the STEC tested, none belonged to the O111 or O113 groups. The amplification with the mPCR-4 assay showed that all isolates harbored the enterohemolysin (ehlyA) gene whereas the intimin marker (eaeA) was observed in all E. coli O157:H7 and O26 isolates.

Słowa kluczowe

PL

zywnosc; mieso wolowe; mieso surowe; zanieczyszczenia bakteryjne; Escherichia coli; szczepy shigatoksyczne; wykrywanie; identyfikacja; sondy molekularne; metoda multiplex PCR; markery patogennosci

• Wydawca: -
• Czasopismo: Medycyna Weterynaryjna
• Rocznik: 2008
• Tom: 64
• Numer: 05
• Opis fizyczny: s.668-672,tab.,bibliogr.

Twórcy

autor

Weiner M

• Panstwowy Instytut Weterynaryjny - Panstwowy Instytut Badawczy, Al.Partyzantow 57, 24-100 Pulawy

Bibliografia

• 1. Al-Gallas N., Ben Aissa R. R., Attia Annabi T., Bahri O., Boudabous A.: Isolation and characterization of Shiga toxin-producing Escherichia coli from meat and dairy products. Food Microbiol. 2002, 19, 389-398.
• 2. Blanco J., Blanco M., Blanco J. E., Mora A., Gonzalez E. A., Bernardez M. I.: Verotoxin-producing Escherichia coli in Spain: Prevalence, serotypes, and virulence genes of O157:H7 and non-O157 VTEC in ruminants, raw beef products, and humans. Exp. Biol. Med. 2003, 228, 345-351.
• 3. Chinen I., Miliwebsky E. S., Chillemi G. M., Baschkier A., Rivas M.: Clonal relatedness of Argentinean Escherichia coli O157:H7 strains by pulsed-field gel electrophoresis. Ninth International Congress Infectious Diseases. Buenos Aires 2000, s. 97.
• 4. Elder R. O., Keen J. E., Siragusa G. R., Barkocy-Gallagher G. A., Koohmaraie M., Laegreid W. W.: Correlation of enterohemorrhagic Escherichia coli O157 prevalence in feces, hides, and carcasses of beef cattle during processing. Proc. Natl Acad. Sci. USA 2000, 97, 2999-3003.
• 5. Khan A., Yamasaki S., Sato T., Ramamurthy T., Pal A., Datta S., Chowdhury N. R., Sas S. C., Sikdar A., Tsukamoto T., Bhattacharya S. K., Takeda Y., Nair G. B.: Prevalence and genetic profiling of virulence determinants of non-O157 Shiga toxin producing Escherichia coli isolated from cattle, beef, and humans, Calcutta, India. Emerg. Infect. Dis. 2002, 8, 54-61.
• 6. Kumar H. S., Otta S. K., Karanasagar I., Karanasagar I.: Detection of Shiga-toxigenic Escherichia coli (STEC) in fresh seafood and meat marketed in Mangalore, India by PCR. Lett. Appl. Microbiol. 2001, 33, 334-338.
• 7. Kwiatek K., Różańska H.: Badania nad występowaniem szczepów enterokrwotocznych Escherichia coli O157:H7 w surowcach i produktach zwierzęcego pochodzenia. XII Konf. Nauk. Diagnostyka mikrobiologiczna. Puławy 19-20.06.1996. Rocz. Wojsk. Inst. Hig. Epidemiol. 1996, 33, 14.
• 8. Meng J., Zhao S., Doyle M. P.: Virulence genes of Shiga toxin-producing Escherichia coli isolated from food, animals and humans. Int. J. Food Microbiol. 1998, 45, 229-235.
• 9. O'Hanlon K. A., Catarame T. M. G., Duffy G., Blair I. S., McDowell D. A.: Rapid detection and quantification of. E. coli O157/O26/O111 in minced beef by real-time PCR. J. Appl. Microbiol. 2004, 96, 1013-1023.
• 10. Osek J.: Aktualne metody diagnostyki szczepów Escherichia coli O157. Medycyna Wet. 2001, 57, 783-787.
• 11. Osek J.: Development of a multiplex PCR approach for the identification of Shiga toxin-producing Escherichia coli strains and their major virulence factor genes. J. Appl. Microbiol. 2003, 95, 1217-1225.
• 12. Osek J., Gallien P.: Zastosowanie specyficznych dla toksyn Shiga sond DNA znakowanych digoksygeniną do oznaczania w kale bydła shigatoksycznych szczepów Escherichia coli (STEC). Medycyna Wet. 2002, 58, 863-866.
• 13. Osek J., Weiner M.: Opracowanie testów multiplex PCR do identyfikacji odmian toksyny Shiga wytwarzanych przez Escherichia coli. Medycyna Wet. 2004, 60, 207-210.
• 14. Paciorek J.: Virulence properties of Escherichia coli faecal strains isolated in Poland from healthy children and strains belonging to serogroups O18, O26, O44, O86, O126 and O127 isolated from children with diarrhoea. J. Med. Microbiol. 2002, 51, 548-556.
• 15. Pierard D., Van Damme L., Moriau L., Stevens D., Lauwers S.: Virulence factors of verocytotoxin-producing Escherichia coli isolated from raw meats. Appl. Environ. Microbiol. 1997, 63, 4585-4587.
• 16. Read S. C., Gyles C. L., Clarke R. C., Lior H., McEwen S.: Prevalence of verocytotoxigenic Escherichia coli in ground beef, pork, and chicken in Southwestern Ontario. Epidemiol. Infect. 1990, 105, 11-20.
• 17. Sadowska B., Osek J., Bonar A., Więćkowska-Szakiel M., Rudnicka W., Różalska B.: Phenotypic and molecular characteristics of typical and atypical Escherichia coli O157, clinical and food isolates. Acta Microbiol. Pol. 2003, 52, 149-158.
• 18. Samadpour M., Kubler M., Buck F. C., Depavia G. A., Mazengia E., Stewart J.: Prevalence of Shiga toxin-producing Escherichia coli in ground beef and cattle feces from King Country, Washington. J. Food Prot. 2002, 65, 1322-1325.
• 19. Szych J., Paciorek J., Cieślik A., Kałużewski S.: Charakterystyka szczepów E. coli O157 izolowanych w Polsce z próbek materiału klinicznego i z żywności. Med. Dośw. Mikrobiol. 1999, 50, 179-196.
• 20. Tarr P. I., Tran T. N., Wilson R. A.: Escherichia coli O157:H7 in retail ground beef in Seattle: results of a one-year perspective study. J. Food Prot. 1999, 62, 133-139.
• 21. Tatarczak M., Wieczorek K., Posse B., Osek J.: Identification of putative adhesin genes in shigatoxigenic Escherichia coli isolated from different sources. Vet. Microbiol. 2005, 110, 77-85.
• 22. Toma C., Espinosa E. M., Song T., Miliwebsky E., Chinen I., Iyoda S., Iwanaga M., Rivas M.: Distribution of putative adhesions in different seropathotypes of Shiga toxin-producing Escherichia coli. J. Clin. Microbiol. 2004, 42, 4937-4946.
• 23. Villari P., Iannuzzo M., Torre I.: An evaluation of the use of 4-methylumbelliferyl-β-D-glucuronide (MUG) in different solid media for the detection and enumeration of Escherichia coli in foods. Lett. Appl. Microbiol. 1997, 24, 286-290.
• 24. Weiner M.: Opracowanie testów multiplex PCR do identyfikacji i charakterystyki shigatoksycznych Escherichia coli. Medycyna Wet. (w druku).