Investigation on the Genetic Signatures of Antibiotic Resistance in Multi-Drug-Resistant Klebsiella Pneumoniae Isolates From National Guard Hospital, Riyadh.

Abstract

IntroductionDespite a large number of antibiotics available to treat Klebsiella (K.) pneumoniae (KP), resistance against these antibiotics is ever-increasing and has now become a global threat to human life. The most frequently observed resistant genes in Klebsiella pneumoniae are CTX-M, OXA-48, IMP, and NDM; some are clone-specific while others form a reservoir for infection.MethodsMatrix-assisted laser desorption ionization-time of flight (MALDI-TOF) was employed for the identification of the pathogens and automated VITEK-2 (bioMérieux, Marcy-l'Étoile, France) was used for minimum inhibitory concentration (MIC) determination, followed by polymerase chain reaction (PCR) amplification of target genes and Sanger sequencing of amplicons.ResultsForty-three out of 50 isolates (86%) were OXA gene-positive, and 49 out of 50 (98%) isolates were CTX-M gene positive. Two phenotypes of OXA were identified in 33 samples sequenced, OXA-505 (70%) and OXA-232 (30%). Sixteen isolates (32%) were positive for NDM-1. Twelve isolates were positive for both OXA and NDM. Multilocus sequence typing (MLST) on these isolates showed that they were distributed in 12 sequence types (STs). Thirty-six out of 50 were grouped in four clonal complexes. ST-14 was the predominant genotype.ConclusionThis study has revealed that CTX-M-15 is the most common extended-spectrum beta-lactamase (ESBL) present in almost all isolates. The study also shows the presence of OXA as the main carbapenemase gene, alone or in combination with other carbapenemases such as NDM-1. Multilocus sequence typing revealed the incidence of polyclonal KP pool with ST-14, ST-29, ST-307, and ST-15 being the predominant ones.