Volume 8, Number 1—January 2002
Research
Prevalence and Genetic Profiling of Virulence Determinants of Non-O157 Shiga Toxin-Producing Escherichia coli Isolated from Cattle, Beef, and Humans, Calcutta, India
Table 1
Polymerase chain reaction (PCR) primers and conditions used in this study
Primer no. | Nucleotide sequence of primers | Target | PCR conditionsa |
||||
---|---|---|---|---|---|---|---|
Denaturing | Annealing | Extension | Amplicon (bp) | Reference | |||
EVT1/ EVT2 | 5'-CAACACTGGATGATCTCAG-3' 5'-CCCCCTCAACTGCTAATA-3' | stx1 family | 94°C, 60s | 55°C, 60sb | 72°C, 60s | 349 | 20 |
EVS1/ EVC2 | 5'-ATCAGTCGTCACTCACTGGT-3' 5'-CTGCTGTCACAGTGACAAA-3' | stx2 family | 94°C, 60s | 55°C, 60sb | 72°C, 60s | 110 | 20 |
hlyA1/ hlyA4 | 5'-GGTGCAGCAGAAAAAGTTGTAG-3' 5'-TCTCGCCTGATAGTGTTTGGTA-3' | EHEC hlyA | 94°C, 30s | 57°C, 60s | 72°C, 90s | 1,551 | 7 |
wkat-B/ wkat-F | 5'-CTTCCTGTTCTGATTCTTCTGG-3' 5'-AACTTATTTCTCGCATCATCC-3' | katP | 94°C, 30s | 56°C, 60s | 72°C, 150s | 2,125 | 9 |
D1/ D13R | 5'-CGTCAGGAGGATGTTCAG-3' 5'-CGACTGCACCTGTTCCTGATTA-3' | etpD | 94°C, 30s | 52°C, 60s | 72°C, 70s | 1,062 | 21 |
EAE1/ EAE2 | 5'-AAACAGGTGAAACTGTTGCC-3' 5'-CTCTGCAGATTAACCTCTGC-3' | eae | 94°C, 60s | 55°C, 90s | 72°C, 90s | 350 | 22 |
a Unless stated, PCR was done for 30 cycles.
b After 35 cycles, final extension step of 10 min at 72°C was performed.
References
- Riley LW, Remis RS, Helgerson SD, McGee HB, Wells JG, Davis BR, Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med. 1983;308:681–5. DOIPubMedGoogle Scholar
- Lopez EL, Contrini MM, De Rosa MF. Epidemiology of Shiga-toxin producing Escherichia coli in South America. In: Kaper JB, O'Brien AD, editors. Escherichia coli and other Shiga-toxin producing E. coli strains. Washington: American Society for Microbiology; 1998. p. 30-7.
- O'Brien AD, Holmes RK. Shiga and Shiga-like toxin. Microbiol Rev. 1987;1:206–20.
- O'Brien AD, Tesh VL, Rolfe AD, Jackson MP, Olsnes S, Sandvig K, Shiga toxin: biochemistry, genetics, mode of action, and role in pathogenesis. Curr Top Microbiol Immunol. 1992;180:65–94. DOIPubMedGoogle Scholar
- Strockbine NA, Marques LR, Newland JW, Smith HW, Holmes RK, O'Brien AD. Two toxin-converting phages from Escherichia coli O157:H7 strain 933 encode antigenically distinct toxins with similar biologic activities. Infect Immun. 1986;53:135–40.PubMedGoogle Scholar
- Kaper JB, Elliott S, Sperandio V, Perna NT, Mayhew GF, Blattner FR. Attaching and effacing intestinal histopathology and the locus of enterocyte effacement. In: Kaper JB, O'Brien AD, editors. Escherichia coli and other Shiga toxin-producing E. coli strains. Washington: American Society for Microbiology; 1998. p. 163-82.
- Schmidt H, Beutin L, Karch H. Molecular analysis of the plasmid-encoded hemolysin of Escherichia coli O157:H7 strain EDL 933. Infect Immun. 1995;63:1055–61.PubMedGoogle Scholar
- Brunder W, Schmidt H, Karch H. KatP, a novel catalase-peroxidase encoded by the large plasmid of enterohaemorrhagic Escherichia coli O157:H7. Microbiology. 1996;142:3305–15. DOIPubMedGoogle Scholar
- Brunder W, Schmidt H, Karch H. EspP, a novel extracellular serine protease of enterohaemorrhagic Escherichia coli O157:H7 cleaves human coagulation factor V. Mol Microbiol. 1997;24:767–78. DOIPubMedGoogle Scholar
- Schmidt H, Henkel B, Karch H. A gene cluster closely related to type II secretion pathway operons of Gram-negative bacteria is isolated on the large plasmid of enterohemorrhagic Escherichia coli O157 strains. FEMS Microbiol Lett. 1997;148:265–72. DOIPubMedGoogle Scholar
- Schmidt H, Geitz C, Phillips IT, Matthias F, Karch H. Non-O157 pathogenic shiga toxin producing Escherichia coli: phenotypic and genetic profiling of virulence traits and evidence for clonality. J Infect Dis. 1999;179:115–23. DOIPubMedGoogle Scholar
- Beutin L, Geier D, Steinruck H, Zimmermann S, Scheutz F. Prevalence and some properties of verotoxin (Shiga-like toxin)-producing Escherichia coli in seven different species of healthy domestic animals. J Clin Microbiol. 1993;31:2483–8.PubMedGoogle Scholar
- Clark RC, Wilson JB, Read SC, Renwick S, Rahn K, Johnson P, Vero cytotoxin producing Escherichia coli (VTEC) in the food chain: preharvest and processing perspectives. In: Karmali MA and Goglio AG, editors. Recent advances in verocytotoxin-producing Escherichia coli infections. Amsterdam: Elsevier Science B.V.; 1994. p. 17-24.
- Hilborn ED, Mermin JH, Mshar PA, Hadler JL, Voetsch A, Wojtkunski C, A multistate outbreak of Escherichia coli O157:H7 infections associated with consumption of mesclun lettuce. Arch Intern Med. 1999;59:1758–64. DOIPubMedGoogle Scholar
- Cody SH, Glynn MK, Farrar JA, Cairns KL, Griffin PM, Kobayashi J, An outbreak of Escherichia coli O157:H7 infection from unpasteurized commercial apple juice. Ann Intern Med. 1999;130:202–9.PubMedGoogle Scholar
- Challmers RM, Aird H, Bolton FJ. Waterborne Escherichia coli O157. J Appl Microbiol. 2000;88:124S–32S.
- Renwick SA, Wilson JB, Clarke RC, Lior H, Borczyk AA, Spika J, Evidence of direct transmission of Escherichia coli O157:H7 infection between calves and a human. J Infect Dis. 1993;168:792–3. DOIPubMedGoogle Scholar
- Gupta S, Soni NK, Kaur P, Sood DK. Verocytopathic activity of Escherichia coli O157 and other “O” serogroups isolated from patients of diarrhea. Indian J Med Res. 1992;95:71–6.PubMedGoogle Scholar
- World Health Organization. Manual for laboratory investigation of acute enteric infections. Geneva: The Organization; 1983.
- Pal A, Ghosh S, Ramamurthy T, Yamasaki S, Tsukamoto T, Bhattacharya SK, Shiga-toxin producing Escherichia coli from healthy cattle in a semi-urban community in Calcutta, India. Indian J Med Res. 1999;110:83–5.PubMedGoogle Scholar
- Schmidt H, Henkel B, Karch H. A gene cluster closely related to type II secretion pathway operons of gram-negative bacteria is located on the large plasmid of enterohemorrhagic Escherichia coli O157 strains. FEMS Microbiol Lett. 1997;148:265–72. DOIPubMedGoogle Scholar
- Yu J, Kaper JB. Cloning and characterization of the eae gene of enterohemorrhagic Escherichia coli O157:H7. Mol Microbiol. 1992;6:411–7. DOIPubMedGoogle Scholar
- Yamasaki S, Lin Z, Shirai H, Terai A, Oku Y, Ito H, Typing of verotoxins by DNA colony hybridization with poly- and oligonucleotide probes, a bead-enzyme-linked immunosorbent assay, and polymerase chain reaction. Microbiol Immunol. 1996;40:345–52.PubMedGoogle Scholar
- Kurazono H, Sasakawa C, Yoshikawa M, Takeda Y. Cloning of a Vero toxin (VT1, Shiga-like toxin 1) gene from a VT1-converting phage isolated from Escherichia coli O157:H7. FEMS Microbiol Lett. 1987;44:23–6. DOIGoogle Scholar
- Ito H, Teral A, Kurazono H, Takeda Y, Nishibuchi M. Cloning and nucleotide sequencing of Vero toxin 2 variant genes from Escherichia coli O91:H21 isolated from a patient with hemolytic uremic syndrome. Microb Pathog. 1990;8:47–60. DOIPubMedGoogle Scholar
- Moseley SL, Huq I, Alim ARMA, So M, Samadpour-Motalebi M, Falkow S. Detection of enterotoxigenic Escherichia coli by DNA colony hybridization. J Infect Dis. 1980;142:892–8. DOIPubMedGoogle Scholar
- Oswald E, Schmidt H, Morabito S, Karch H, Marches O, Caprioli A. Typing of intimin genes in human and animal enterohemorrhagic and enteropathogenic Escherichia coli: Characterization of a new intimin variant. Infect Immun. 2000;68:64–71. DOIPubMedGoogle Scholar
- Bhan M, Raj P, Levine MM, Kaper JB, Bhandari N, Srivastava R, Enteroagggregative Escherichia coli associated with persistent diarrhoea in a cohort of rural children in India. J Infect Dis. 1989;159:1061–4. DOIPubMedGoogle Scholar
- Albert MJ, Faruque SM, Faruque ASG, Neogi PKB, Ansaruzzaman M, Bhuiyan NA, Controlled study of Escherichia coli diarrheal infections in Bangladeshi children. J Clin Microbiol. 1995;33:973–7.PubMedGoogle Scholar
- Germani Y, Cunin P, Tedjouka E. bou Ncharre C, Morvan J, Martin P. Enterohaemorrhagic Escherichia coli in Ngoila (Cameroon) during an outbreak of bloody diarrhoea. Lancet. 1998;352:625–6. DOIPubMedGoogle Scholar
- World Health Organization. Zoonotic non-O157 shiga toxin-producing Escherichia coli (STEC). Report of a WHO Scientific Working Group Meeting. WHO/CSR/APH/98.8. Geneva: The Organization; 1998. http://www.who.int/emc.
- Radu S, Mutalib SA, Rusel G, Ahmed Z, Morigaki T, Asea N, Detection of Escherichia coli O157:H7 in the beef marketed in Malaysia. Appl Environ Microbiol. 1998;64:1153–5.PubMedGoogle Scholar
- Karmali MA, Martin P, Lim C, Fleming PC, Arbus GS, Lior H. The association between the idiopathic hemolytic uremic syndrome and infection by verotoxin-producing Escherichia coli. J Infect Dis. 1985;151:775–82. DOIPubMedGoogle Scholar
- Karmali MA, Martin P, Lim C, Cheung R, Arbus GS. Sensitive method for detecting low numbers of verotoxin-producing Escherichia coli in mixed cultures by use of colony sweeps and polymyxin extraction of verotoxin. J Clin Microbiol. 1985;22:614–9.PubMedGoogle Scholar