2017 ◽  
Vol 4 (suppl_1) ◽  
pp. S685-S685
Author(s):  
Patrick Reich ◽  
Mary Boyle ◽  
Melanie Sullivan ◽  
Patrick G Hogan ◽  
Ryley Thompson ◽  
...  

mBio ◽  
2019 ◽  
Vol 10 (4) ◽  
Author(s):  
Santosh K. Bikkarolla ◽  
Viveka Nordberg ◽  
Fredrika Rajer ◽  
Vilhelm Müller ◽  
Muhammad Humaun Kabir ◽  
...  

ABSTRACTThe global spread of antibiotic resistance amongEnterobacteriaceaeis largely due to multidrug resistance plasmids that can transfer between different bacterial strains and species. Horizontal gene transfer of resistance plasmids can complicate hospital outbreaks and cause problems in epidemiological tracing, since tracing is usually based on bacterial clonality. We have developed a method, based on optical DNA mapping combined with Cas9-assisted identification of resistance genes, which is used here to characterize plasmids during an extended-spectrum β-lactamase (ESBL)-producingEnterobacteriaceaeoutbreak at a Swedish neonatal intensive care unit. The outbreak included 17 neonates initially colonized with ESBL-producingKlebsiella pneumoniae(ESBL-KP), some of which were found to carry additional ESBL-producingEscherichia coli(ESBL-EC) in follow-up samples. We demonstrate that all ESBL-KP isolates contained two plasmids with theblaCTX-M-15gene located on the smaller one (~80 kbp). The same ESBL-KP clone was present in follow-up samples for up to 2 years in some patients, and the plasmid carrying theblaCTX-M-15gene was stable throughout this time period. However, extensive genetic rearrangements within the second plasmid were observed in the optical DNA maps for several of the ESBL-KP isolates. Optical mapping also demonstrated that even though other bacterial clones and species carryingblaCTX-Mgroup 1 genes were found in some neonates, no transfer of resistance plasmids had occurred. The data instead pointed toward unrelated acquisition of ESBL-producingEnterobacteriaceae(EPE). In addition to revealing important information about the specific outbreak, the method presented is a promising tool for surveillance and infection control in clinical settings.IMPORTANCEThis study presents how a novel method, based on visualizing single plasmids using sequence-specific fluorescent labeling, could be used to analyze the genetic dynamics of an outbreak of resistant bacteria in a neonatal intensive care unit at a Swedish hospital. Plasmids are a central reason for the rapid global spread of bacterial resistance to antibiotics. In a single experimental procedure, this method replaces many traditional plasmid analysis techniques that together provide limited details and are slow to perform. The method is much faster than long-read whole-genome sequencing and offers direct genetic comparison of patient samples. We could conclude that no transfer of resistance plasmids had occurred between different bacteria during the outbreak and that secondary cases of ESBL-producingEnterobacteriaceaecarriage were instead likely due to influx of new strains. We believe that the method offers potential in improving surveillance and infection control of resistant bacteria in hospitals.


2010 ◽  
Vol 59 (6) ◽  
pp. 687-692 ◽  
Author(s):  
Ali A. Dashti ◽  
Mehrez M. Jadaon ◽  
Huda H. Gomaa ◽  
Bobby Noronha ◽  
Edet E. Udo

The spread of antibiotic-resistant bacteria has become a large problem in most countries including Kuwait. This antibiotic resistance is usually due to the production of extended-spectrum β-lactamase (ESBL) enzymes such as SHV, TEM and CTX-M. This study reports the emergence and spread of an ESBL-producing Klebsiella pneumoniae clone in a neonatal intensive care unit (NICU) in a Kuwaiti hospital. Eight ESBL-producing K. pneumoniae isolates were from blood cultures of seven neonates, and two were from the fingers of two healthcare workers in a NICU in Al Jahra Hospital, Kuwait. All isolates were obtained in February–March 2006, except for one, which was obtained in August 2005. Identification of the bacteria was based on traditional bacteriological and biochemical tests using the Vitek system. Antibiotic susceptibility was tested by the disc diffusion method using 16 different antibiotics. ESBLs were detected using disc approximation and double-disc synergy methods and confirmed as ESBLs using Etest. PCR and DNA sequencing were performed to determine the genotypes and mutations in the β-lactamase genes (bla TEM, bla SHV and bla CTX-M). Genetic relatedness was determined by PFGE. All isolates were confirmed to have ESBLs by the Vitek system, disc approximation test, double-disc diffusion test and Etest, being resistant to cefotaxime, ceftazidime, cefepime, gentamicin, tobramycin and ciprofloxacin but susceptible to tetracycline and trimethoprim–sulfamethoxazole. Molecular studies showed the isolates to have TEM-1 β-lactamase, a CTX-M-15-like ESBL and the newly discovered SHV-112 ESBL. PFGE showed that all isolates had identical banding patterns. The results indicate that a single clone of ESBL-producing K. pneumoniae caused bloodstream infections among babies in a NICU of a Kuwaiti hospital, and may have emerged at least 5 years ago. This clone was also present on the hands of healthcare workers, suggesting that they may have been involved in its transmission. Further studies are recommended to determine whether this clone is also spreading in other Kuwaiti hospitals.


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