Genes
The Campylobacter jejuni/coli MLST scheme uses internal fragments of the following seven house-keeping genes:
aspA (aspartase)
atpA (ATP synthase alpha subunit)
glnA (glutamine synthetase)
gltA (citrate synthase)
glyA (serine hydroxy methyl transferase)
pgm (phospho glucomutase)
tkt (transketolase)
PCR Amplification and sequencing
The primer pairs used for the PCR amplification and sequencing are detailed below:
aspA
Forward
aspAF1: GAGAGAAAAGCWGAAGAATTTAAAGAT
Reverse
aspAR1: TTTTTTCATTWGCRSTAATACCATC
atpA
Forward
atpAF: GWCAAGGDGTTATYTGTATWTATGTTGC
Reverse
atpAR: TTTAADAVYTCAACCATTCTTTGTCC
glnA
Forward
glnAF: TGATAGGMACTTGGCAYCATATYAC
Reverse
glnAR: ARRCTCATATGMACATGCATACCA
gltA
Forward
gltAF: GARTGGCTTGCKGAAAAYAARCTTT
Reverse
gltAR: TATAAACCCTATGYCCAAAGCCCAT
glyA
Forward
glyAF: ATTCAGGTTCTCAAGCTAATCAAGG
Reverse
glyAR: GCTAAATCYGCATCTTTKCCRCTAAA
pgm
Forward
pgmF1: CATTGCGTGTDGTTTTAGATGTVGC
Reverse
pgmR1: AATTTTCHGTBCCAGAATAGCGAAA
tkt
Forward
tktFN: AACGGTTTTAAGTTATCAT
Reverse
tktRN: TTTTGTGACTTCCTTCAAGC
Reaction conditions - PCR
Denaturation: 94ºC 30 s
Annealing: 53ºC 30 s
Extension: 72ºC 2 minutes
30 cycles