TABLE 5.
List of primer and probe sequences used in the multiplex PCRa
| Name | No. of bases | Position in genome | Sequence (5′ to 3′)b |
|---|---|---|---|
| Primers | |||
| invA forward | 20 | 2924483–2924506 | AGCGTACTGGAAAGGGAAAG |
| invA reverse | 24 | 2924598–2924579 | ATACCGCCAATAAAGTTCACAAAG |
| prt forward | 22 | 2175083–2175104 | AGCTCCATAGAAATGCTCCAAT |
| prt reverse | 22 | 2175213–2175192 | GAACATCACTGCCACCAAATAC |
| tyv forward | 25 | 2174357–2174376 | ACTAAGTATATGCCTGATAGCTGTT |
| tyv reverse | 20 | 2174454–2174462 | GCCGTACTGCCTCAAGTAAA |
| Sdf-1 forward | 26 | 1472650–1472675 | CTTTCTCAGATTCAGGGAGTATATCA |
| Sdf-1 reverse | 23 | 1472772–1472750 | TGAACTACGTTCGTTCTTCTGGT |
| IAC forward | 27 | 16–42 | CTGATCTGACTTCACTCCTGTCTACTA |
| IAC reverse | 24 | 116–93 | GACACTCGTCACTGACCACTATCA |
| Probes | |||
| invA (+) | 26 | 2924529–2924554 | 6FAM/CGTCACCTTTGATAAACTTCATCGCA–BHQ1 |
| prt (−) | 26 | 2175150–2175125 | Cy3/CCGCCGCCATTATAGATAAAGTTTGT–BHQ2 |
| tyv (−) | 25 | 2174454–2174430 | Cy3/TGCAGGTCAAGTGGCAATGACTACA–BHQ2 |
| Sdf-1 (+) | 24 | 1472690–1472713 | TexRdXN/ATCAGCCTGTTGTCTGCTCACCAT–BHQ2 |
| IAC (+) | 28 | 54–81 | Cy5/CGCTGACTCGTCCATACAATAAGACACC–BHQ2 |
a
The sequences, lengths, and locations of primers and probes are shown within the Salmonella enterica serovar Enteritidis genome (strain SA20082034, accession no. CP007425.2). The sequence, length and location of the IAC primers and probe are shown on the IAC gene. The lengths of the PCR products generated by the primer pairs were as follows: invA, 115 nucleotides; IAC, 100 nucleotides; prt, 130 nucleotides; tyv, 130 nucleotides; and Sdf-1, 123 nucleotides.
b
6FAM, 6-carboxyfluorescein; BHQ1, black hole quencher 1; TexRd, Texas Red.