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
Asis Khan*, Shinji Yamasaki*†, Toshio Sato†, Thandavarayan Ramamurthy*, Amit Pal*, Simanti Datta*, Nandini Roy Chowdhury*, Suresh Chandra Das‡, Asim Sikdar‡, Teizo Tsukamoto§, Sujit Kumar Bhattacharya*, Yoshifumi Takeda¶, and Gopinath Balakrish Nair*#
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.
