ISO 9001: We are pleased to announce that the BIGSdb-Pasteur platform has obtained ISO9001 certification in March 2026, and has been included in the scope of the CRBIP ISO 9001:2015 certification.
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 |