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Primers used for MLST of Bordetella
Genes
The Bordetella MLST scheme uses internal fragments of the following seven housekeeping genes:
adk (Adenylate kinase)
fumC (fumarate hydratase class II)
glyA (serine hydroxymethyltransferase)
tyrB (aromatic amino-acid aminotransferase)
icd (isocitrate dehyrogenase)
pepA (cytosol aminopeptidase)
pgm (phosphoglucomutase)
PCR amplification
The PCR conditions were optimized using HotStar Taq mastermix (Qiagen) and 1M betaine final concentration. The cycling conditions were as follows:
- 1 cycle:
- 95°C for 15 min
- 30 cycles:
- 95°C for 30 s
- 54°C for 30 s
- 1 cycle:
- 72°C for 1 min
Allele templates
| Gene | Direction | Primer Sequence |
|---|---|---|
| adk | F | AGCCGCCTTTCTCACCCAACACT |
| R | TGGGCCCAGGACGAGTAGT | |
| fumC | F | CGTGAACCGGGGCCAGTCGTC |
| R | GGCCAGCCAGCGCACATCGTT | |
| glyA | F | CAACCAGGGCGTGTACATGGC |
| R | CCGCGATGACGTGCATCAG | |
| tyrB | F | CGAGACCTACGCTTATTACGAT |
| R | TGCCGGCCAGTTCATTTT | |
| icd | F | CTGGTCCACAAGGGCAACAT |
| R | ACACCTGGGTGGCGCCTTC | |
| pepA | F | CGCCCCAGGTTGAAGAAAATCGTC |
| R | ATCAGGCCCACCACATCCAG | |
| pgm | F | CGCCCATGTCACCAGCACCGA |
| R | CGCCGTCTATCGTAACCAG |