MLST of Listeria monocytogenes

The first description of this scheme can be found in Salcedo et al. (2003). Development of a multilocus sequence typing method for analysis of Listeria monocytogenes clones. J Clin Microbiol 41:757-62.

Genes

The Listeria monocytogenes MLST scheme uses fragments of the following seven house-keeping genes:

ABC transporter (abcZ)
Beta-glucosidase (bglA)
Catalase (cat)
Succinyl diaminopimelate dessucinylase (dapE)
D-amino acid aminotransferase (dat)
L-lactate dehydrogenase (ldh)
Histidine kinase (lhkA)

PCR

Bacterial cell lysates were obtained by sonication (10 min) followed by centrifugation for 5 min. The PCR conditions were initial denaturation at 94°C for 4 min followed by 25 cycles of denaturation at 94°C for 30 s, annealing at 52°C for 30 s (except for bglA, which had an annealing temperature of 45°C), and extension at 72°C for 2 min followed by a final extension step of 72°C for 10 min.

Primers used for PCR amplification and sequencing