Volume 13, Number 8—August 2007
Dispatch
PCR versus Hybridization for Detecting Virulence Genes of Enterohemorrhagic Escherichia coli
Robert S. Gerrish*, James E. Lee*, June Reed*, Joel Williams*, Larry D. Farrell*, Kathleen M. Spiegel*, Peter P. Sheridan*, and Malcolm S. Shields*
Table
Virulence factor targets and primers, including nucleotide sequences, reference, and PCR conditions*
| Primer name | Nucleotide sequence (5′→3′) | Target (bp) | Ref. | PCR conditions |
||
|---|---|---|---|---|---|---|
| Denature | Anneal | Extension | ||||
| STX1U | GTAACATCGCTCTTGCCACA | Stx1 gene (204) | This study | 95°C, 60 s | 53.7°C, 60 s | 72°C, 240 s |
| STX1D | CGCTTTGCTGATTTTTCACA | |||||
| STX2U | GTTCCGGAATGCAAATCAGT | Stx2 gene (206) | This study | 95°C, 60 s | 53.7°C, 60 s | 72°C, 240 s |
| STX2D | CGGCGTCATCGTATACACAG | |||||
| eae-1 | ACGTTGCAGCATGGGTAACTC | Intimin (818) | (8) | 95°C, 60 s | 57.1°C, 60 s | 72°C, 240 s |
| eae-2 | GATCGGCAACAGTTTCACCTG | |||||
| ToxBF | TGGCCTTGCGCTCTATAAGAACCT | ToxB (823) | This study | 95°C, 60 s | 60°C, 60 s | 72°C, 240 s |
| ToxBR | ACCACGCCGTGAGAATAATGTCCA | |||||
| HlyA1F | GGTGCAGCAGAAAAAGTTGTAG | HlyA (1551) | (13) | 95°C, 60 s | 55.5°C,60 s | 72°C, 240 s |
| HlyA1R | TCTCGCCTGATAGTGTTTGGTA | |||||
| EspPF | CGGCAGAGTATCATCAAGAGC | EspP (397) | This study | 95°C, 60 s | 55.5°C, 60 s | 72°C, 240 s |
| EspPR | CATTAAATGGAGTTATGCGTC | |||||
| KatPF | TTTAAAACGCTGGGATTTGC | KatP (1174) | This study | 95°C,60 s | 52.0°C, 60 s | 72°C, 240 s |
| KatPR | CTCCTGAGAGGCGTCAGTTC | |||||
| MalBU | GACCTCGGTTTAGTTCACAGA | MalB promoter (414) | This study | 95°C, 60 s | 55.8°C, 60 s | 72°C, 240 s |
| MalBDn | AGCGCGTAGGACTGAAACACCATA | |||||
| ORF1F | TTTTTCAAAGCAAATGATGTGG | ORF 1 pOSAK1 (385) | This study | 95°C, 60 s | 49.8°C, 60 s | 72°C, 240 s |
| ORF1R | GGCGTAGCTAGGTTGAAATTATG | |||||
| ORF2F | CAA CCTAGCTACGCCACCAT | ORF 2 pOSAK1 (869) | This study | 95°C, 60 s | 54.3°C, 60 s | 72°C, 240 s |
| ORF2R | CATCAGGCGGAAATACCACT | |||||
| EAF1 | CAGGGTAAAAGAAAGATGATAA | Eaf (397) | (14) | 95°C, 60 s | 49.8°C, 60 s | 72°C, 240 s |
| EAF2 | TATGGGGACCATGTATTATCA | |||||
| BFP1 | GATTGAATCTGCAATGGC | Bfp (597) | (15) | 95°C, 60 s | 51.6°C, 60 s | 72°C, 240 s |
| BFP2 | GGATTACTGTCCTCACATAT | |||||
*Ref., reference; ORF, open reading frame.
