ABSTRACT
Abstract Escherichia coli is one of the main human pathogens causing different hospital- and community-acquired infections. During the period from January 2013 to March 2015, 1.96% (32/1632) of E. coli isolates recovered at the Hospital Regional de Ushuaia, Tierra del Fuego province, were resistant to third-generation cephalosporins (TGCs). These isolates were resistant to cefotaxime (91%) and/or ceftazidime (28%). No resistance to carbapenems was detected. Twenty-six isolates were positive for blaCTX-M gene, grouped as CTX-M-1/15 (54%); CTX-M-9/14 (25%); CTX-M-2 (17%); and CTX-M-1/15 plus CTX-M-9/14 (4%). Five TGC-resistant strains were positive for blaCMY gene, while one strain harbored TEM-19 ESBL. Twelve isolates were identified as ST131 E. coli hyperepidemic clone, and one as ST69. Genome sequence analysis of seven blaCTX-M-15 E. coli selected isolates confirm the circulation of ST131, ST617 and ST405 international high-risk clones in the city of Ushuaia.
Resumen Escherichia coli es uno de los principales patógenos humanos causantes de diferentes infecciones de inicio hospitalario y comunitario. Se determinó que el 1,96% (32/1.632) de los aislamientos de E. coli recuperados entre enero de 2013 y marzo de 2015 en el Hospital Regional de Ushuaia, provincia de Tierra del Fuego, fueron resistentes a cefalosporinas de tercera generación (CTG). Estos aislamientos fueron resistentes a cefotaxima (91%) y/o a ceftazidima (28%). No se detectó resistencia a los carbapenemes. Veintiséis aislamientos fueron positivos para el gen blaCTX-M, agrupados como CTX-M-1/15 (54%), CTX-M-9/14 (25%), CTX-M-2 (17%) y CTX-M-1/15 más CTX-M-9/14 (4%). Cinco cepas resistentes a CTG dieron positivo para el gen blaCMY, mientras que un aislamiento presentó la BLEE TEM-19. Doce aislamientos se identificaron como clon hiperepidémico E. coli ST131 y uno como ST69. El análisis de las secuencias del genoma de siete aislamientos seleccionados de E. coli blaCTX-M-15 confirmó la circulación de los clones internacionales de alto riesgo ST131, ST617 y ST405 en la ciudad de Ushuaia.
ABSTRACT
Escherichia coli can harbor genomic pks islands that code for a polyketide-peptide genotoxin known as colibactin. E. coli strains carrying pks islands trigger genetic instability. pks islands have been significantly associated with bacteremia. We investigated the molecular epidemiology of bacteremic E. coli isolates and the prevalence of bacteremia-causing E. coli carrying pks islands. A total of 146 E. coli isolates were collected at a tertiary-care hospital from January 2015 to December 2016. The phylogenetic groups were determined by multiplex PCR. All isolates were screened by PCR for sequence type 131 (ST131)-associated single-nucleotide polymorphisms (SNPs) in mdh and gyrB. For detection of pks islands, we performed PCR for the clbB and clbN genes as colibactin system markers. Phylogenetic group B2 was the most common, accounting for 54.1% (N=79) of the isolates, followed by group D with 29.5% (N=43), group A with 11.6% (N=17), and group B1 with 4.8%. Of the group B2 isolates, 40.5% were ST131 strains and 32.9% carried pks islands. Only three ST131 isolates in group B2 carried the clbB and clbN genes, while the other 23 ST131 isolates did not. The pks gene might not be associated with ST131 strains.
Subject(s)
Humans , Bacteremia , Escherichia coli , Escherichia , Islands , Molecular Epidemiology , Multiplex Polymerase Chain Reaction , Polymerase Chain Reaction , PrevalenceABSTRACT
OBJECTIVE@#To investigate the phylogenetics and prevalence of bloodstream infections with ST131, the antimicrobial resistance profiles of the pathogens, and the clinical features.@*METHODS@#Non-duplicate isolates were collected from 144 patients with bloodstream infections in our hospital between January and December, 2016.The phylogenetic groups of the isolates were analyzed using multiplex PCR, and O serotyping of ST131 strains was performed by allele-specific PCR.The clinical characteristics of the 144 patients were analyzed to define the differences in the clinical features between patients with ST131 infection and those with non-ST131 infection.Antibiotic susceptibility of the isolates was determined using the Vitek 2 compact system.@*RESULTS@#The phylogenetic group analysis showed a domination by group B2 (41.0%[59/144]), followed by group F, group B1 and group E, which accounted for 16.7%(24/144), 13.9%(20/144), and 13.2% (19/144), respectively.Nine strains (6.3%) of were identified to be ST131 strains, among which 8 were O25b-B2-ST131 strains and 1 was O16-B2-ST131 strain.Of the 9 cases of ST131 infection, 7(77.8%) were found to occur in a nosocomial setting.The demographic characteristics and clinical features of the ST131-infected patients were similar to those of non-ST131-infected patients.ST131 strains were sensitive to piperacillin/tazobactam, imipenem, ertapenem, and amikacin, but showed high resistance rates to cefazolin, ceftriaxone, ciprofloxacin, levofloxacin, gentamicin, and trimethoprim/ sulfamethoxazole (all over 50%).The positivity rate of ESBLs in the ST131 strains was 77.8%, and the multidrug resistance rate reached 88.9%, which was higher than that of non-ST131 isolates, but the difference was not statistically significant.@*CONCLUSIONS@#The most common phylogenetic groups of isolates from patients with bloodstream infections are group B2 and F, and the positivity rate of ST131 is low.We for the first time detected O16-ST131 in patients with blood-borne infections in China.The clinical features of ST131-infected patients are similar to those of non-ST131-infected patients.The positivity rate of ESBLs and the multidrug resistance rate are high in ST131 strains, which may raise concerns in the future.
Subject(s)
Humans , Anti-Bacterial Agents , Therapeutic Uses , Bacteremia , Drug Therapy , Epidemiology , Microbiology , China , Drug Resistance, Bacterial , Escherichia coli , Classification , Genetics , Escherichia coli Infections , Drug Therapy , Epidemiology , Microbiology , Genotype , Microbial Sensitivity Tests , Molecular Epidemiology , Phylogeny , Species SpecificityABSTRACT
Objective To investigate the molecular epidemiology and antimicrobial resistance status of uropathogenic Escherichia coli (UPEC) in senior population in Putuo District ,Shanghai .Methods A total of 72 UPEC strains were isolated from elderly inpatients with urinary tract infections in Putuo Hospital ,Shanghai University of Traditional Chinese Medicine from January 2013 to March 2015 .The strains were characterized by multi‐locus sequence typing (MLST ) .The β‐lactamase gene and the plasmid mediated quinolone resistance (PMQR) gene were detected ,and the mutations of quinolone resistance‐determining regions (QRDR) in gyrA and parC genes were demonstrated .In vitro drug susceptibility test was performed .Continuous variables were compared using t test and categorical variables were compared using chi‐squared test or Fisher exact test .Results The UPEC strains showed different resistance rates to ciprofloxacin ,cefotaxime and trimethoprim‐sulfamethoxazole ,which were 76 .4% ,73 .6% and 65 .3% , respectively .UPEC still remained highly sensitive to imipenem ,meropenem ,amikacin and piperacillin‐tazobactam .Among 72 isolates ,55 (76 .4% ) of 49 (68 .1% ) extended‐spectrum β‐lactamase (ESBL )‐positive strains harbored blaCTX‐M genes .Among the 55 ciprofloxacin resistant strains ,51 (92 .7% ) had three or four mutations in QRDR of gyrA and parC genes .The “hot‐spot” mutations of QRDR were located at amino acid position 83 and 87 in gyrA gene and at positions 80 and 84 in parC gene .Forward analysis by MLST showed that the most frequent sequence types (ST ) were ST131 (18/72 ,25 .0% ) , ST1193(7/72 ,9 .7% ) ,ST405 (7/72 ,9 .7% ) ,ST38 (5/72 ,6 .9% ) and ST648 (3/72 ,4 .2% ) .ST131 isolates were predominant in ST which caused community‐onset urinary tract infections .Multiple drug‐resistance were detected in ST 131 ,ST405 ,ST38 and ST648 which were mainly producing blaCTX‐M ESBL .Conclusions Community‐acquired multiple drug‐resistant UPEC strains such as ST131 clone are prevalent in elderly patients .Thus ,monitoring of molecular epidemiology would be beneficial to prevent the prevalence of multiple drug‐resistant UPEC .
ABSTRACT
Background: Extended-spectrum β-lactamase (ESBL) producing uropathogens has become prevalent worldwide. E. coli O25b-ST131 clone, associated with blaCTX-M-15, has been reported from many parts of the world and is frequently associated with multidrug resistance. Thus far, there are no reports about this clone in Bangladesh. The objective of this study was to investigate ESBL producing uropathogens and to survey the prevalence of E. coli O25b-ST131 clone among ESBL positive E. coli isolates. Methods: From symptomatic urinary tract infection cases, a total of 800 urine samples were collected. Bacterial identification and antimicrobial susceptibility testing was performed using established methods. Screening of ESBL producers was done using the disk diffusion method. Screening positive isolates were phenotypically confirmed by double disk synergy (DDS) test. Genes encoding ESBLs (blaCTX-M-15, blaOXA-1) were identified both by PCR and DNA sequencing. Phenotypic positive ESBL producers were also studied by PCR for existence of class 1 integron. Subsequently, O25b-ST131 clone was identified by allele specific PCR. Results: Of 138 gram-negative uropathogens, 45 (32.6%) were positive for ESBLs. ESBL producers showed high frequency of antimicrobial resistance except imipenem. Among 45 ESBL producers, 36 (80%) produced blaCTX-M-15, 18 (40%) produced blaOXA-1. Fifteen (33.3%) strains simultaneously produced both blaOXA-1 and blaCTX-M-15. Class 1 integron was present in 30 (66.7%) isolates. Of the 31 blaCTX-M-15 positive E. coli, 22 (71%) were positive for E. coli O25b-ST131 clone and all (100%) belonged to B2 phylogenetic group. Conclusion: Rising antimicrobial resistance among uropathogens, and especially the emergence of blaCTX-M-15 positive E. coli O25b-ST131 clone in Bangladesh has provided urgency to the development of novel preventive and therapeutic strategies.
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Objective To investigate the molecular types of carbapenem-non-susceptible Esche-richia coli ( E.coli) isolates and their mechanism of carbapenem resistance .Methods Twenty-two carbap-enem-non-susceptible E.coli strains were isolated from 3 hospitals in Hangzhou from 2007 to 2011.The mini-mum inhibitory concentrations ( MICs) of antimicrobials to those isolates were determined by agar dilution method and E-test.The molecular mechanisms of carbapenem resistance of E.coli isolates were analyzed by conjugation experiment,PCR and DNA sequencing.Pulsed-field gel electrophoresis (PFGE),multilocus se-quence typing ( MLST ) , and phylogenetic typing were performed to analyze the molecular epidemiology of those isolates.Results The MICs of imipenem and meropenem to 22 E.coli isolates were ranged from 1 μg/ml to 16 μg/ml,and the MICs of ertapenem were 2 μg/ml to 64 μg/ml.All E.coli isolates produced the KPC-2 carbapenemase and various β-lactamases , and some of them also produced plasmid-mediated AmpC enzymes.Carbapenem resistance was transferred by conjugation and transformation from 22 E.coli iso-lates to E.coli EC600 strains.The E.coli transconjugants or transformants acquired the blaKPC-2 gene and showed similar antibiotic susceptibility patterns in comparison with donor strains .Only a few isolates were in-distinguishable or closely related as indicated by PFGE .Four sequence types including ST131 (9 isolates), ST648 (5 isolates),ST38 (2 isolates) and ST405 (2 isolates) were identified by MLST.Phylogenetic analy-sis indicated that 9 ST131 isolates belonged to phylogenetic group B 2 and the other isolates belonged to group D (11 isolates),group B1 (1 isolate) and group A (1 isolate),respectively.Conclusion The sequence type of prevalent E.coli isolates producing KPC-2 from Hangzhou was ST131,which is an international epi-demic,multidrug-resistant clone,followed by ST648.