Objective To isolate a strain of Listeria monocytogenes from frozen chicken meat sold in Gansu Province and analyze its serotype, antibiotic resistance phenotype, and genomic characteristics. Methods The slide agglutination method was used for serological identification, the broth microdilution method was used to analyze the sensitivity of the strain to 8 kinds of antibiotics, and the genomic characteristics were analyzed by sequencing. Results The strain was identified as Listeria monocytogenes by 16S rDNA species identification, with a 1/2a serotype, phylogenetic lineage II, subline SL 155, and clone CC 155 MLST 705, cgMLST 13240. Sensitive to 4 kinds of antibiotics including ampicillin, penicillin, vancomycin, and meropenem, and multidrug-resistant to 4 kinds of antibiotics including erythromycin, ciprofloxacin, tetracycline, and compound sulfamethoxazole. Simultaneously carrying aph(3')-III, ant(6)-Ia, dfrG, fosX, lnu(B), lsa(E), erm(B), tet(S) and tet(M) 9 kinds of resistance genes, carrying 5 virulence genes of LIPI-1, prfA gene deletion, carrying 12 kinds of internalization genes, no premature stop codon of inlA gene carrying, no LIPI-3, and LIPI-4. Conclusions The Listeria monocytogenes carries abundant virulence genes and resistance genes, and its resistance phenotype is basically consistent with the resistance genes, showing multiple drug resistance, providing a reference for the prevention and treatment of listeriosis.