MLST Pasteur Web Site






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Primers used for MLST of Listeria monocytogenes

- Genes

The Listeria monocytogenes MLST scheme uses internal fragments of the following seven housekeeping genes:

abcZ (ABC transporter)

bglA (beta glucosidase)

cat (catalase)

dapE (succinyl diaminopimelate desuccinylase)

dat (D-amino acid aminotransferase)

ldh (L-lactate dehydrogenase)

lhkA (histidine kinase)

- PCR amplification

Protocol uses primers with universal sequencing tails to sequence all genes with the same forward and reverse sequencing primers.

abcZoF : GTTTTCCCAGTCACGACGTTGTATCGCTGCTGCCACTTTTATCCA

abcZoR : TTGTGAGCGGATAACAATTTCTCAAGGTCGCCGTTTAGAG

bglAoF : GTTTTCCCAGTCACGACGTTGTAGCCGACTTTTTATGGGGTGGAG

bglAoR : TTGTGAGCGGATAACAATTTCCGATTAAATACGGTGCGGACATA

catoF : GTTTTCCCAGTCACGACGTTGTAATTGGCGCATTTTGATAGAGA

catoR : TTGTGAGCGGATAACAATTTCAGATTGACGATTCCTGCTTTTG

dapEoF : GTTTTCCCAGTCACGACGTTGTACGACTAATGGGCATGAAGAACAAG

dapEoR : TTGTGAGCGGATAACAATTTCATCGAACTATGGGCATTTTTACC

datoF : GTTTTCCCAGTCACGACGTTGTAGAAAGAGAAGATGCCACAGTTGA

datoR : TTGTGAGCGGATAACAATTTCTGCGTCCATAATACACCATCTTT

ldhoF : GTTTTCCCAGTCACGACGTTGTAGTATGATTGACATAGATAAAGA

ldhoR : TTGTGAGCGGATAACAATTTCTATAAATGTCGTTCATACCAT

lhkAoF : GTTTTCCCAGTCACGACGTTGTAAGAATGCCAACGACGAAACC

lhkAoR : TTGTGAGCGGATAACAATTTCTGGGAAACATCAGCAATAAAC

For strains of lineage 3, we use the following primers in case of failure with the above primers:

lhkA-F3 : GCAAGTTTTGAATACGTATCAGCG

lhkA-R2 : TACGCATTTCATGAGAAACATCAG

- Universal sequencing primers

oF : GTT TTC CCA GTC ACG ACG TTG TA

oR : TTG TGA GCG GAT AAC AAT TTC

- PCR conditions

94° for 4 min

94° for 30 s

52° for 30 s* (35 cycles)

72° for 2 min

72° for 10 min

*PCR amplification is performed at an annealing temperature of 52°C for all genes except for bglA (45°C).

- Allele templates

abcZ(537 bp)

AAATCGACGAACAGAATGCGTATAGGACTTTTCCGCAAGATGGAAAAACTATCAATCCGTTTCTTCGATAGTCGCAATGATGG

CGAAATGCTTAGTCGTTTCACTAGTGACTTAGATAATATTTCCAACACACTAAACCAAGCATTGATCCAAGTACTATCCAACG

TCGCGCTAATGATTGGTGTTATCATCATGATGTTCCAACAAAACGTGGAACTAGCCTTCGTTACTCTAATATCTGCTCCATTT

GCGATTATTATTGCGACAGTGATTATTCGAAAAGCCCGCAAATTCGTTGATATTCAACAAGATGAACTAGGCGTACTTAACGG

CTACATTGACGAAAAAATCTCTGGTCAAAAAATTATTATCACAAATGGCTTAGAAGAAGAAACAATTGACGGCTTTGTTAAAC

AAAACAATATCGTTAAAAACGCAACTTACAAAGGTCAAGTTTACTCCGGTTTACTTTTCCCAATGATGCAAGGTATTTCCCTA

TTAAATACAGCTATTGTTATCTTCTTCGGTGGATGGTTA

bglA (399 bp)

AACCAATTCGAAGGCGCTTACAACGTCGATGGAAAAGGACTTTCCGTTCAAGATGTTACTCCAAAAGGCGGATTTGGTCACAT

TACTGACGGTCCAACACCAGATAACTTAAAATTAGAAGGAATTGACTTCTATCATCGCTACAAAGATGACGTGAAACTTTTTG

CCGAAATGGGCTTCAAGGTTTTCCGTACTTCCATCGCTTGGTCCCGTATCTTCCCAAATGGTGACGAAACTGAGCCAAACGAA

GCAGGATTACAATTTTACGATGATTTATTCGATGAACTTCTAGCACATAATATCGAACCACTGATTACTTTATCTCACTATGA

AACACCACTTCACTTATCGAAAACTTACGACGGCTGGGTAAATAGAAAAATGATCGACTTCTATGAA

cat (486 bp)

GCTCGTGGTGCTGGTGCTCACGGGAAATTTGTCACTAAGAAAAGCATGAAAAAATATACAATGGCTAAATTTTTGCAAGAAGA

AGGAACGGAAACGGAGGTTTTTGCTCGTTTTTCAACAGTAATTCATGGGCAACATTCTCCAGAAACATTACGTGATCCACGCG

GTTTCTCCGTTAAGTTTTATACAGAAGAAGGGAATTATGATTTTGTCGGAAATAATTTGCCGGTATTCTTCATTCGTGATGCG

ATTAAGTTTCCGGATGTTATTCATTCCTTGAAGCCTGATCCACGCACAAATATTCAAGATGGCAACCGTTACTGGGATTTCTT

TAGCCTTACACCGGAAGCTACGACGATGATTATGTACTTATTCAGTGATGAAGGAACGCCGGCTTCTTACCGGGAAATACGTG

GCTCTAGTGTTCATGCGTTCAAATGGATTAATGAAGAAGGCAAAACAGTTTATGTAAAACTGCGCTGGATT

dapE (462 bp)

TTGCAAAAGTTGTTAGCTGAACACGGTATTGAGTCCGAAAAGGTAAAATACGACGTAGACAGAGCCAGCCTAGTTAGCGAAAT

TGGTTCCAGTGACGAGAAAGTTTTGGCGTTTTCAGGGCATATGGATGTCGTTGATGCGGGTGATGTCTCGAAGTGGAAGTTCC

CACCTTTTGAAGCAACAGAGCATGAAGGGAAAATATACGGACGTGGCGCGACGGATATGAAGTCAGGTCTAGCGGCGATGATT

ATTGCAATGATTGAGCTTCATGAAGAAAAACAAAAACTAAATGGCAAAATTAGATTATTAGCAACGGTTGGTGAAGAAGTCGG

TGAACTTGGAGCCGAACAACTAACGCAAAAAGGTTACGCAGATGATTTAGATGGCTTGATTATCGGCGAACCGAGTGGACACC

GGATTGTTTATGCGCATAAAGGTTCCATTAATTATACCGTTAAATCC

dat (471 bp)

GAAGTAGTTCGTCTATATAATGGAAAATTCTTCACTTATAATGAACACATTGATCGTTTATATGCGAGTGCAGCAAAAATTGA

CTTAGTTATTCCTTATTCTAAAGAAGAGTTACGAGCGTTACTTGAAAAATTAGTTGCTGAAAATAATATTAATACAGGAAATG

TCTATTTACAAGTGACTCGAGGTGTTCAAAACCCGCGTAATCACGTTATGCCAGATGATTTCCCGCTGGAAGGCGTTTTAACA

GCAGCAGCTCGTGAAGTACCAAGAAATGAACAACAATTTGTGCAAGGTGGACCAGTAATTACAGAAGAAGATGTTCGTTGGTT

ACGCTGTGACATCAAGAGTTTGAATTTACTTGGAAACATTTTAGCAAAAAACAAAGCACATCAACAAAATGCGTTAGAAGCTG

TTTTACACCGCGGAGAGCAAGTAACTGAGTGTTCAGCTTCCAATATTTCTATTATT

ldh (453 bp)

TATAGCGACTGCCACGATGCGGACTTAGTTGTTGTAACTGCCGGGACTGCACAAAAACCTGGTGAAACTCGTTTAGATTTAGT

AAATCGTAATATTAAAATCATGAAAGGCATCGTGGATGAAGTAATGGCTAGCGGATTTGACGGTATCTTCTTAATCGCTTCTA

ACCCAGTAGATATCTTAACTTACGCTACATGGAAATTCTCAGGTCTTCCAAAAGAACGTGTTATCGGTTCTGGAACAAGCCTT

GATACAGCACGTTTCCGTATGTCAATTGCCGACTATCTAAAAGTAGATGCTCGTAACGTCCATGGTTACATCCTTGGCGAACA

CGGCGATACAGAATTCCCAGCATGGAGCCACACAACTGTCGGCGGTCTTCCAATCACTGAATGGATTAGCGAAGATGAACAAG

GTGCAATGGATACTATTTTCGTAAGTGTTCGTGATGCA

lhkA (480 bp)

TATCCAACACAGATGAATCAGCCGTTACCAAAGGATTTCTCTATTTCTTCGGATGATAAGAAAAAACTTGAAAGTGGCGAAAC

AGTTAGTAAGAAAATAGATAATCGCTTTAATAAAGAAATGACAATTGTGTACGTCCCAATAATGAATGGCGACAAATTTGTCG

GTTCTATCGTGCTCAATTCACCTATTAGCGGTACGGAGCAAGTAATTGGTACGATTAACCGCTATATGTTCTACACTATTTTA

CTTTCTATAACGGTAGCACTTATTCTTAGCGCAATCTTGTCCAAACTACAAGTAAATCGAATCAACAAACTACGAGCAGCGAC

AAAAGACGTTATTCAAGGCAATTACAAAGCTCGATTGAAGGAAAATAATTTTGATGAAATTGGTGCACTTGCCATTGATTTCA

ATAAAATGACACAAACCCTTGAAACATCTCAAGAAGAAATAGAACGACAAGAGAAGCGGAGACGC



Databases are maintained by Sylvain Brisse using software developed by Keith Jolley at Oxford (Jolley et al. 2004, BMC Bioinformatics, 5:86). Web managers : Ghislaine GUIGON and Jean-michel THIBERGE This site is hosted by the technological platform Genotyping of Pathogens and Public Health* of Institut Pasteur.*