Listeria monocytogenes is an important foodborne pathogen. The aims of this study were to determine genetic relatedness of L. monocytogenes isolated from ready-to-eat (RTE) foods in Malaysia. L monocytogenes isolates from RTE foods were characterized by multiplex-PCR serotyping, REP-PCR, BOX-PCR, RAPD, PFGE, virulotyping and antibiotyping. Of the 32 L. monocytogenes isolates analyzed, 21 (65.6%) were assigned to serogroup "1/2a, 3a", seven (21.9%) serogroup "1/2c, 3c", and four (12.5%) serogroup "4h, 4d, 4e". All the L. monocytogenes harbored InlA, inlB, inlC and inlJ virulence genes. More than half (53%) L. monocytogenes isolates were resistant to penicillin G, followed by tetracycline (15.6%), amoxicillin-clavulanic acid (12.5%), vancomycin (9.4%) erythromycin (6.3%), clindamycin, streptomycin, kanamycin, and chloramphenicol (each 3.1%). REP-PCR, BOX-PCR, RAPD and PFGE generated 28 (D = 0.992), 31 (D = 0.998), 32 (D = 1), and 20 (D = 0.916) patterns, respectively. These results indicate that L monocytogenes isolates from RTE food were heterogeneous. There was no correlation between antibiograms and serogroups or pulsotypes or PCR-typing and/or sources of isolates. Since different subtyping methods often give different discriminatory powers, the use of more than one subtyping approach is necessary in providing a more accurate picture of the genetic diversity of L. monocytogenes. In conclusion, L monocytogenes isolates from RTE possess the internalin genes and are genetically diverse. Furthermore, the occurrence of resistant isolates belonging to epidemiologically important serogroups "1/2a, 3a" and "4h, 4d, 4e" in RTE foods is a matter of public health concern. (C) 2014 Elsevier Ltd. All rights reserved.