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The outbreak of carbapenem-resistant Klebsiella pneumoniae is a serious public health problem, especially in the neonatal intensive care unit (NICU).Fifteen K. pneumoniae strains were isolated from 7 neonates during June 3 to 28, 2017 in an NICU. Antimicrobial susceptibility was determined by the Vitek 2 system and microbroth dilution method. Multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) were used to analyze the genetic relatedness of the isolates. Whole-genome sequencing and gene function analysis were performed to investigate pathogenicity and drug resistance and screen genomic islands.Three clones of K. pneumoniae were identified from 7 neonates: 7 strains of ST37, 7 of novel ST3006, and 1 of ST1224. Gene sequencing showed that the kpn1343 (ST37) strain harbored 12 resistance genes (OXA-33, TEM-1, SHV-11, AAC (6’)-IId, AAC (3)-IIa, AAC (6’)-Ib-cr, catB3, arr-3, sul1, oqxB, oqxA, CRP, and catB3) and included 15 genomic islands and 205 reduced virulence genes. The kpn1344 (ST3006) strain harbored 4 antibiotic-resistant genes (TEM-1, CTX-M-3, vgaC, and CRP) and included 19 genomic islands and 209 reduced virulence genes. MLST and PFGE showed that 15 strains of K. pneumoniae were divided into 3 groups with a high level of homology. ST1224 (kpn1362) was isolated on June 28, 2017, which was 10 days after the last isolate (kpn1359, June 18, 2017); thus, we speculated that ST1224 was not the clone that caused the outbreak.This co-outbreak of K. pneumoniae involved 2 clones: ST37 and ST3006. ST37 carried the multidrug-resistant genes, such as OXA-33, TEM-1, and SHV-11, and ST3006 was a novel K. pneumoniae ST typing. Whole-genome sequencing may be an effective method for screening bacterial-resistant genes and their functions.