Genes
The neisserial MLST scheme uses internal fragments of the
following seven house-keeping genes:
abcZ (putative ABC transporter)
adk (adenylate kinase)
aroE (shikimate dehydrogenase)
fumC (fumarate hydratase)
gdh (glucose-6-phosphate dehydrogenase)
pdhC (pyruvate dehydrogenase subunit)
pgm (phosphoglucomutase)
PCR Amplification
Please note: These are new primer
sequences (added 10 October 2003). Amplification primers, with the
exception of those for pgm, were designed by A. Birtles at the Meningococcal
Reference Unit, Health Protection Agency, UK.
The primer pairs we use for the PCR amplification of internal
fragments of these genes are:
abcZ-P1C 5-TGTTCCGCTTCGACTGCCAAC-3
abcZ-P2C 5-TCCCCGTCGTAAAAAACAATC-3
adk-P1B 5-CCAAGCCGTGTAGAATCGTAAACC-3
adk-P2B 5-TGCCCAATGCGCCCAATAC-3
aroE-P1B 5-TTTGAAACAGGCGGTTGCGG-3
aroE-P2B 5-CAGCGGTAATCCAGTGCGAC-3
fumC-P1B 5-TCCCCGCCGTAAAAGCCCTG-3
fumC-P2B 5-GCCCGTCAGCAAGCCCAAC-3
gdh-P1B 5-CTGCCCCCGGGGTTTTCATCT-3
gdh-P2B 5-TGTTGCGCGTTATTTCAAAGAAGG-3
pdhC-P1B 5-CCGGCCGTACGACGCTGAAC-3
pdhC-P2B 5-GATGTCGGAATGGGGCAAACA-3
pgm-P1 5'-CTTCAAAGCCTACGACATCCG-3'
pgm-P2 5'-CGGATTGCTTTCGATGACGGC-3'
Sequencing
We use the following primer pairs for sequencing:
abcZ-S1A (P1A) 5'-AATCGTTTATGTACCGCAGR-3'
abcZ-S2 5'-GAGAACGAGCCGGGATAGGA-3'
adk-S1A 5'-AGGCWGGCACGCCCTTGG-3'
adk-S2 5'-CAATACTTCGGCTTTCACGG-3'
aroE-S1A 5-GCGGTCAAYACGCTGRTK-3
aroE-S2 5-ATGATGTTGCCGTACACATA-3
fumC-S1 5-TCCGGCTTGCCGTTTGTCAG-3
fumC-S2 5-TTGTAGGCGGTTTTGGCGAC-3
gdh-S3 5'-CCTTGGCAAAGAAAGCCTGC-3'
gdh-S4C 5-RCGCACGGATTCATRYGG-3
pdhC-S1 5'-TCTACTACATCACCCTGATG-3'
pdhC-S2 5'-ATCGGCTTTGATGCCGTATTT-3'
pgm-S1 5'-CGGCGATGCCGACCGCTTGG-3'
pgm-S2A 5'-GGTGATGATTTCGGTYGCRCC-3'