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2.
Rev. Soc. Bras. Med. Trop ; 53: e20200064, 2020. tab, graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS, SES-SP | ID: biblio-1136871

ABSTRACT

Abstract INTRODUCTION: Carbapenem-resistant Klebsiella pneumoniae infection lacks treatment options and is associated with prolonged hospital stays and high mortality rates. The production of carbapenemases is one of the most important factors responsible for this multi-resistance phenomenon. METHODS: In the present study, we analyzed the presence of genes encoding carbapenemases in K. pneumoniae isolates circulating in one of the public hospitals in the city of Aracaju, Sergipe, Brazil. We also determined the best combination of drugs that display in vitro antimicrobial synergy. First, 147 carbapenem-resistant K. pneumoniae isolates were validated for the presence of blaKPC, bla GES, bla NDM, bla SPM, bla IMP, bla VIM, and bla OXA-48 genes using multiplex polymerase chain reaction. Thereafter, using two isolates (97 and 102), the role of double and triple combinational drug therapy as a treatment option was analyzed. RESULTS: Seventy-four (50.3%) isolates were positive for bla NDM, eight (5.4%) for bla KPC, and one (1.2%) for both bla NDM and bla KPC. In the synergy tests, double combinations were better than triple combinations. Polymyxin B and amikacin for isolate 97 and polymyxin B coupled with meropenem for isolate 102 showed the best response. CONCLUSIONS: Clinicians in normal practice use multiple drugs to treat infections caused by multi-resistant microorganism; however, in most cases, the benefit of the combinations is unknown. In vitro synergistic tests, such as those described herein, are important as they might help select an appropriate multi-drug antibiotic therapy and a correct dosage, ultimately reducing toxicities and the development of antibiotic resistance.


Subject(s)
Humans , Bacterial Proteins/genetics , beta-Lactamases/genetics , Klebsiella Infections/microbiology , Klebsiella pneumoniae/drug effects , Anti-Bacterial Agents/pharmacology , Brazil , DNA, Bacterial , Microbial Sensitivity Tests , Polymerase Chain Reaction , Prevalence , Drug Resistance, Multiple, Bacterial/genetics , Klebsiella pneumoniae/isolation & purification , Klebsiella pneumoniae/genetics
3.
Rev. Soc. Bras. Med. Trop ; 53: e20190526, 2020. tab, graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS, SES-SP | ID: biblio-1136834

ABSTRACT

Abstract INTRODUCTION: This study investigated the genetic environment of bla KPC-2 in Klebsiella pnemoniae multi-drug resistant clinical isolates. METHODS: Four carbapenemase gene isolates resistant to carbapenems, collected from infected patients from two hospitals in Brazil, were investigated using polymerase chain reaction and plasmid DNA sequencing. RESULTS: The bla KPC-2 gene was located between ISKpn6 and a resolvase tnpR in the non-Tn4401 element (NTEKPC-IId). It was detected on a plasmid belonging to the IncQ1 group. CONCLUSIONS To our knowledge, this is the first report of the presence of the bla KPC-2 gene in the NTEKPC-IId element carried by plasmid IncQ1 from infections in Brazil.


Subject(s)
Humans , beta-Lactamases/genetics , Klebsiella Infections/microbiology , Klebsiella pneumoniae/genetics , Anti-Bacterial Agents/pharmacology , Plasmids/genetics , DNA, Bacterial/genetics , Microbial Sensitivity Tests , Polymerase Chain Reaction , Drug Resistance, Multiple, Bacterial , Klebsiella pneumoniae/isolation & purification , Klebsiella pneumoniae/enzymology
5.
Rev. Assoc. Med. Bras. (1992) ; 65(5): 678-681, May 2019. graf
Article in English | LILACS | ID: biblio-1012972

ABSTRACT

SUMMARY OBJECTIVE: We describe the case of a diabetic patient who developed vertebral osteomyelitis and bilateral psoas abscess with gas formation due to klebsiella pneumoniae. METHODS: A 64-year-old woman with a 4-year history of type-2 diabetes mellitus was admitted to the Emergency Department. The subject had a 2-day history of high-grade fever associated with chills and a 5-hour history of consciousness. She received empirical treatment with febrifuge, after which her fever decreased. RESULTS: Her fever recurred after an interval of three hours. A computed tomography scan of the abdomen revealed vertebral osteomyelitis and bilateral psoas muscle abscess with gas formation. Blood culture and purulent fluid described the growth of the Klebsiella pneumoniae. The patient received antibiotic therapy and bilateral drainage therapy after the drainage catheter was placed into the abscess cavity by CT-guidance. Due to the serious damage to the vertebral column and permanent pain, the patient underwent minimally invasive internal spinal fixation and recovered successfully. CONCLUSION: A case of vertebral osteomyelitis and bilateral psoas abscess with gas formation caused by Klebsiella pneumoniae in a diabetic patient. Antibiotic therapy, drainage, and minimally invasive internal spinal fixation were performed, which enabled a good outcome.


RESUMO OBJETIVO: Descrever o caso de uma paciente diabética que desenvolveu osteomielite vertebral e abcesso bilateral do psoas com formação de gás causada por klebsiella pneumoniae. MÉTODOS: Uma mulher de 64 anos de idade, com 4 anos de histórico de diabetes mellitus tipo 2, foi admitida no Serviço de Emergência. A paciente apresentava um quadro de dias de febre alta acompanhada de calafrios e um histórico de 5 horas de consciência. Ela recebeu tratamento empírico com antitérmico, após o qual a febre diminuiu. RESULTADOS: A febre retornou após um intervalo de três horas. Uma tomografia computadorizada do abdome revelou osteomielite vertebral e abcesso bilateral do músculo psoas com formação de gás. A cultura do sangue e o fluido purulento revelaram o crescimento de Klebsiella pneumoniae. A paciente recebeu antibióticos e terapia de drenagem bilateral após o cateter de drenagem ser posicionado na cavidade do abscesso com auxílio de TC. Devido a sérios danos à coluna vertebral e a dor permanente, a paciente foi submetida à fixação vertebral interna minimamente invasiva e recuperou-se com sucesso. CONCLUSÃO: Um caso de osteomielite vertebral e abscesso do psoas bilateral com a formação de gás causada por Klebsiella pneumoniae em uma paciente diabética. Antibioticoterapia, drenagem e fixação vertebral interna minimamente invasiva foram realizadas, o que permitiu um bom resultado.


Subject(s)
Humans , Female , Osteomyelitis/surgery , Spinal Diseases/surgery , Klebsiella Infections/surgery , Psoas Abscess/surgery , Diabetes Complications/surgery , Klebsiella pneumoniae/pathogenicity , Osteomyelitis/microbiology , Spinal Diseases/microbiology , Klebsiella Infections/microbiology , Tomography, X-Ray Computed/methods , Drainage/methods , Reproducibility of Results , Treatment Outcome , Psoas Abscess/microbiology , Diabetes Complications/microbiology , Gases/metabolism , Middle Aged
6.
Einstein (Säo Paulo) ; 17(4): eGS4444, 2019. tab
Article in English | LILACS | ID: biblio-1001916

ABSTRACT

ABSTRACT Objective: To estimate the direct medical costs of drug therapy of Klebsiella pneumoniae carbapenemase (KPC) infection patients in hospital-based context. Methods: A cost-of-illness study conducted with a prospective cohort design with hospitalized adults infected by KPC. Data collection was performed using an instrument composed of sociodemographic data, clinical and prescription medication. Estimates of the direct costs associated to each treatment were derived from the payer's perspective, in the case of federal public hospitals from Brazil, and included only drug costs. These costs were based on the average price available at the Brazilian Price Database Health. No discount rate was used for the cost of drugs. The costs are calculate in American Dollar (US$). Results: A total of 120 inpatients participated of this study. The total drug cost of these inpatients was US$ 367,680.85. The systemic antimicrobial group was responsible for 59.5% of total costs. The direct drug cost per patients infected by KPC was conservatively estimated at nearly US$ 4,100.00, and about of 60% of costs occurred during the period of infection. Conclusion: The findings of our study indicate a thoughtful economic hazard posed by KPC that all healthcare sectors have to face. The increasing worldwide incidence of these bacteria represents a growing burden that most health systems are unable to deal with. There is an imperative need to develop protocols and new antimicrobials to treatment of KPC, aiming to rearrange resources to increase the effectiveness of healthcare services.


RESUMO Objetivo: Estimar os custos médicos diretos da terapia medicamentosa de pacientes com infecção por carbapenemase por Klebsiella pneumoniae carbapenemase (KPC) em contexto hospitalar. Métodos: Estudo de custo de doença realizado com desenho de coorte prospectiva, com adultos hospitalizados infectados por KPC. A coleta de dados foi realizada usando instrumento composto por dados sociodemográficos, medicamentos clínicos e prescritos. As estimativas dos custos diretos associados a cada tratamento foram derivadas da perspectiva dos pagadores, no caso dos hospitais públicos federais do Brasil, e incluíram apenas custos de medicamentos, os quais basearam-se no preço médio disponível na Price Database Health do Brasil. Nenhuma taxa de desconto foi utilizada para o custo dos medicamentos. Os custos foram calculados em dólares norte-americanos (US$). Resultados: Um total de 120 pacientes hospitalizados participou do estudo. O custo total da droga desses pacientes internados foi de US$ 367,680.85. O grupo antimicrobianos de uso sistêmico foi responsável por 59,5% dos custos totais. O custo direto estimado de forma conservadora, por paciente, foi de aproximadamente US$ 4,100.00, e cerca de 60% destes se deram durante o período de infecção. Conclusão: Os achados deste estudo apontam um risco econômico importante relacionado a KPC, o qual todos os setores de saúde terão que enfrentar. A incidência mundial em elevação destas bactérias representa carga crescente, e a maioria dos sistemas de saúde é incapaz de resolvê-la. Há necessidade imperativa de se desenvolverem protocolos e novos antimicrobianos para o tratamento de KPC, com o objetivo de reorganizar os recursos para aumentar a efetividade dos serviços de saúde.


Subject(s)
Humans , Male , Female , beta-Lactamases , Klebsiella Infections/economics , Prospective Studies , Hospitalization/economics , Klebsiella pneumoniae/isolation & purification , Klebsiella pneumoniae/enzymology , Anti-Bacterial Agents/economics , Bacterial Proteins , Klebsiella Infections/microbiology , Klebsiella Infections/drug therapy , Health Care Costs , Hospitalization/statistics & numerical data , Inpatients , Middle Aged , Anti-Bacterial Agents/administration & dosage
7.
Braz. j. microbiol ; 49(4): 885-890, Oct.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-974312

ABSTRACT

ABSTRACT In this study, the performance of the "RESIST-3 O.K.N. K-SeT" (Coris BioConcept, Gembloux, Belgium) immunochromatographic assay was evaluated in 132 Klebsiella pneumoniae comprising 102 carbapenem resistant and 30 carbapenem susceptible isolates. Genotypically known isolates of Gram negative bacteria (n = 22) including various species were also tested by the assay as controls. The isolates tested by the immunochromatographic assay and also were run PCR for bla KPC, bla IMP, bla VIM, bla NDM, and bla OXA-48. The rates of bla NDM, bla OXA-48, and bla KPC in carbapenem resistant isolates were found at 52.9%, 39.2%, and 2.0%, respectively. Both bla NDM and bla OXA-48 were found in six (5.9%) isolates. The results of the assay showed 100% concordance with those obtained by PCR in 132 K. pneumoniae. The agreement between the two methods was found to be identical at the isolate level. The assay also correctly detected all genotypically known isolates of Escherichia coli, Serratia marcescens, Citrobacter freundii, Enterobacter cloacae, K. pneumoniae carrying bla KPC, bla NDM, and/or bla OXA-48. On the other hand, the assay did not exhibit any cross-reaction in control isolates harboring bla IMP and bla VIM. We conclude that the RESIST-3 O.K.N. K-SeT is a reliable, rapid, and user friendly test and we recommend it for routine diagnostic laboratories.


Subject(s)
Humans , Bacterial Proteins/analysis , beta-Lactamases/analysis , Klebsiella Infections/microbiology , Immunoassay/methods , Klebsiella pneumoniae/enzymology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Turkey , beta-Lactamases/metabolism , Carbapenems/pharmacology , Polymerase Chain Reaction , Klebsiella pneumoniae/isolation & purification , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/chemistry , Anti-Bacterial Agents/pharmacology
8.
Salud pública Méx ; 60(1): 29-40, Jan.-Feb. 2018. tab, graf
Article in English | LILACS | ID: biblio-903844

ABSTRACT

Abstract: Objective: To compare the genetic determinants involved in plant colonization or virulence in the reported genomes of K. variicola, K. quasipneumoniae and K. pneumoniae. Materials and methods: In silico comparisons and Jaccard analysis of genomic data were used. Fimbrial genes were detected by PCR. Biological assays were performed with plant and clinical isolates. Results: Plant colonization genes such as cellulases, catalases and hemagglutinins were mainly present in K. variicola genomes. Chromosomal β-lactamases were characteristic of this species and had been previously misclassified. K. variicola and K. pneumoniae isolates produced plant hormones. Conclusions: A mosaic distribution of different virulence- and plant-associated genes was found in K. variicola and in K. quasipneumoniae genomes. Some plant colonizing genes were found mainly in K. variicola genomes. The term plantanosis is proposed for plant-borne human infections.


Resumen: Objetivo: Comparar genes de colonización de plantas o de virulencia en los genomas reportados de K. variicola, K. quasipneumoniae y K. pneumoniae. Material y métodos: Se utilizaron análisis in silico y de Jaccard. Por PCR se detectaron genes de fimbrias. Se realizaron ensayos biológicos con aislados de plantas y clínicos. Resultados: Los genes de colonización de plantas como celulasas, catalasas y hemaglutininas se encontraron principalmente en genomas de K. variicola. Las β-lactamasas cromosómicas son características de la especie y en algunos casos estaban mal clasificadas. K. variicola y K. pneumoniae producen hormonas vegetales. Conclusiones: Se encontró una distribución en mosaico de los genes de asociación con plantas y de virulencia en K. variicola y K. quasipneumoniae. Principalmente en K. variicola se encontraron algunos genes involucrados en la colonización de plantas. Se propone el término plantanosis para las infecciones humanas de origen vegetal.


Subject(s)
Humans , Plants/microbiology , Klebsiella Infections/microbiology , Klebsiella/physiology , Bacterial Proteins/physiology , Bacterial Proteins/genetics , Virulence/genetics , Computer Simulation , Disease Reservoirs , Adaptation, Biological/genetics , Genome, Bacterial , Drug Resistance, Multiple, Bacterial , Gene Ontology , Genes, Bacterial , Klebsiella/enzymology , Klebsiella/genetics , Klebsiella/pathogenicity
9.
Salud pública Méx ; 60(1): 56-62, Jan.-Feb. 2018. tab, graf
Article in English | LILACS | ID: biblio-903842

ABSTRACT

Abstract: Objective: Due to the fact that K. variicola, K. quasipneumoniae and K. pneumoniae are closely related bacterial species, misclassification can occur due to mistakes either in normal biochemical tests or during submission to public databases. The objective of this work was to identify K. variicola and K. quasipneumoniae genomes misclassified in GenBank database. Materials and methods: Both rpoB phylogenies and average nucleotide identity (ANI) were used to identify a significant number of misclassified Klebsiella spp. genomes. Results: Here we report an update of K. variicola and K. Quasipneumoniae genomes correctly classified and a list of isolated genomes obtained from humans, plants, animals and insects, described originally as K. pneumoniae or K. variicola, but known now to be misclassified. Conclusions: This work contributes to recognize the extensive presence of K. variicola and K. quasipneumoniae isolates in diverse sites and samples.


Resumen: Objetivo: Identificar genomas mal clasificados de K. variicola, y K. quasipneumoniae en la base de datos del GenBank. Material y métodos: En el presente estudio se usaron tanto análisis filogenéticos usando rpoB como la identidad media de nucleótidos (ANI, por sus siglas en ingles) para identificar un número significativo de genomas del género Klebsiella. Resultados: Se reportó una actualización de genomas de K. variicola y K. quasipneumoniae correctamente clasificados y una lista de aquellos aislamientos obtenidos de seres humanos, plantas, animales e insectos, descritos originalmente como K. pneumoniae o K. variicola pero ahora se conoce que están mal clasificados. Conclusiones: Este trabajo contribuye a la presencia extensiva de aislamientos de K. variicola y K. quasipneumoniae en diversos sitios y muestras.


Subject(s)
Animals , Plants/microbiology , Ursidae/microbiology , Klebsiella Infections/microbiology , Bacterial Typing Techniques , Genome, Bacterial , Insecta/microbiology , Klebsiella/classification , Phylogeny , DNA, Bacterial , Sequence Analysis, DNA
10.
Braz. j. microbiol ; 49(supl.1): 93-100, 2018. tab, graf
Article in English | LILACS | ID: biblio-974332

ABSTRACT

Abstract Klebsiella pneumoniae is important human and animal pathogen that causes a wide spectrum of infections. In this study, isolates from cattle nasal swabs samples were identified by 16S rRNA, and to evaluate the antimicrobial susceptibility, virulence gene carrying levels, and multilocus sequence typing of K. pneumoniae isolates. 33 isolates of K. pneumoniae were isolated and identified in 213 nasal swabs samples, of which 12 were hypervirulent K. pneumoniae strains. Extended Spectrum Beta-Lactamases genes were found in 93.4% of the strains. Of which, TEM was the most prevalent (93.4%), followed by CTX-M and SHV were 57.6% and 39.4%, respectively. A main mutation pattern of quinoloneresistance-determining region, Thr83-Ieu and Asp87-Asn in gyrA and Ser87-Ile in parC, was detected in 33 K. pneumoniae isolates. All the isolates harbored at least two virulence factor genes, with ureA (97.0%) and wabG (91.0%) exhibiting high carriage rates in 33 K. pneumoniae isolates. MLST revealed 7 sequence types, of which 3 STs (2541, 2581 and 2844) were newly assigned. Using eBURST, ST2844 and ST2541 were assigned to new clonal complex 2844. Our study provides evidence and biological characteristics of K. pneumoniae isolates from cattle upper respiratory tract in Southwest China.


Subject(s)
Animals , Cattle , Bacterial Proteins/genetics , Klebsiella Infections/veterinary , Cattle Diseases/microbiology , Drug Resistance, Multiple, Bacterial , Virulence Factors/genetics , Klebsiella pneumoniae/isolation & purification , Klebsiella pneumoniae/drug effects , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/metabolism , Klebsiella Infections/microbiology , China , Virulence Factors/metabolism , Multilocus Sequence Typing , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/metabolism
11.
Rev. chil. infectol ; 35(3): 239-245, 2018. tab, graf
Article in Spanish | LILACS | ID: biblio-959437

ABSTRACT

Resumen Introducción: La emergencia de Klebsiella productora de carbapenemasas resistente a colistín representa un desafío clínico y un problema emergente. Objetivo: Evaluar la mortalidad intrahospitalaria y sus potenciales factores de riesgo en pacientes internados con infecciones clínicas por Klebsiella pneumoniae productora de carbapenemasas (KPC) resistente a colistín. Material y Método: Realizamos un estudio de cohorte retrospectivo, incluyendo pacientes adultos admitidos a un hospital universitario de tercer nivel en Buenos Aires, infectados por KPC resistente a colistín. El evento primario considerado fue la mortalidad intrahospitalaria. Se utilizaron modelos generalizados lineales para evaluar potenciales predictores de dicho evento. Resultados: En total, se identificaron 18 pacientes hospitalizados que presentaron una infección clínica por esta bacteria durante el año 2016 y que fueron incluidos en el análisis final. La mortalidad intrahospitalaria en esta cohorte fue de 38,9%. La presencia de bacteriemia, la injuria renal aguda al momento del diagnóstico y la presencia de shock séptico se asociaron a la ocurrencia del evento primario. Conclusión: El desarrollo de infecciones clínicamente relevantes por KPC resistente a colistín en pacientes internados es frecuente y presenta una elevada mortalidad. En nuestra cohorte, la presencia de shock e injuria renal aguda al momento del diagnóstico se asociaron a un incrementado riesgo de mortalidad intrahospitalaria. Futuras investigaciones deberían corroborar estos hallazgos e investigar factores adicionales que permitan identificar tempranamente a aquellos pacientes que presentarán eventos desfavorables.


ABSTRACT Background: The emergence of colistin resistant carbapenemase-producing Klebsiella represents a therapeutic challenge and a worldwide problem. Aim: To estimate the in-hospital mortality and identify the associated risk factors among patients with colistin-resistant carbapenemase-producing Klebsiella pneumoniae (KPC) that present with a clinical infection. Methods: We carried a retrospective cohort study, including adult patients infected with colistin-resistant KPC hospitalized at a tertiary teaching hospital in Buenos Aires, Argentina during the year 2016. The main outcome was in-hospital mortality. We used generalized lineal models to evaluate potential predictors of mortality. Results: 18 patients that developed a colistin-resistant KPC clinical infection were identified and included in the final analysis. In-hospital mortality in this cohort was 38.9%. The presence of bacteremia, acute renal injury at the time of diagnosis and septic shock were associated with the main outcome. Conclusions: Infections due to colistin-resistant KPC among in-hospital patients was frequent and was associated with high mortality rate. In our cohort, both shock and acute kidney injury were associated with a higher likelihood of poor outcomes. Further studies are warranted to evaluate the role of these and others risk factors so as to aid in the early detection of high risk patients.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Bacterial Proteins/metabolism , beta-Lactamases/metabolism , Klebsiella Infections/mortality , Hospital Mortality , Colistin , Klebsiella pneumoniae/drug effects , Anti-Bacterial Agents/therapeutic use , Argentina , Klebsiella Infections/enzymology , Klebsiella Infections/microbiology , Klebsiella Infections/drug therapy , Retrospective Studies , Risk Factors , Drug Resistance, Bacterial
12.
Medwave ; 18(2): e7191, 2018.
Article in English, Spanish | LILACS | ID: biblio-912086

ABSTRACT

Las carbapenemasas son uno de los mecanismos enzimáticos de resistencia antimicrobiana, que compromete la mayor parte de los antibióticos betalactámicos. Por lo general, su producción se debe al uso indiscriminado de antimicrobianos. A nivel mundial, la expansión de este mecanismo de resistencia es inminente y las medidas de control son limitadas. Con el objeto de discutir los problemas relacionados a este mecanismo emergente de resistencia, reportamos un caso de Klebsiella pneumoniae productora de carbapenemasas en Huancayo, la Región de la Sierra Central de Perú.


Carbapenemases are one of the major mechanisms of antimicrobial resistance, usually due to the indiscriminate use of antibiotics. The expansion of this mechanism of resistance at world level is imminent and control measures are limited. In the region of the Central Sierra of Peru - Huancayo, we report a case of carbapenemase-producing Klebsiella pneumoniae, with the purpose of discussing the problems related to this emerging mechanism of antibiotic resistance.


Subject(s)
Humans , Male , Adult , Klebsiella Infections/drug therapy , Klebsiella pneumoniae/drug effects , Anti-Bacterial Agents/pharmacology , Peru , Bacterial Proteins/metabolism , beta-Lactamases/metabolism , Klebsiella Infections/microbiology , Drug Resistance, Bacterial , Klebsiella pneumoniae/isolation & purification , Klebsiella pneumoniae/enzymology
14.
Braz. j. microbiol ; 48(3): 509-514, July-Sept. 2017. tab
Article in English | LILACS | ID: biblio-889143

ABSTRACT

Abstract The production of KPC (Klebsiella pneumoniae carbapenemase) is the major mechanism of resistance to carbapenem agents in enterobacterias. In this context, forty KPC-producing Enterobacter spp. clinical isolates were studied. It was evaluated the activity of antimicrobial agents: polymyxin B, tigecycline, ertapenem, imipenem and meropenem, and was performed a comparison of the methodologies used to determine the susceptibility: broth microdilution, Etest® (bioMérieux), Vitek 2® automated system (bioMérieux) and disc diffusion. It was calculated the minimum inhibitory concentration (MIC) for each antimicrobial and polymyxin B showed the lowest concentrations for broth microdilution. Errors also were calculated among the techniques, tigecycline and ertapenem were the antibiotics with the largest and the lower number of discrepancies, respectively. Moreover, Vitek 2® automated system was the method most similar compared to the broth microdilution. Therefore, is important to evaluate the performance of new methods in comparison to the reference method, broth microdilution.


Subject(s)
Humans , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/metabolism , beta-Lactamases/metabolism , Enterobacteriaceae Infections/microbiology , Klebsiella Infections/microbiology , Klebsiella pneumoniae/drug effects , Microbial Sensitivity Tests/methods , Bacterial Proteins/genetics , beta-Lactamases/genetics , beta-Lactams/pharmacology , Drug Resistance, Bacterial , Enterobacter/drug effects , Enterobacter/genetics , Enterobacter/isolation & purification , Klebsiella pneumoniae/classification , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/isolation & purification , Polymyxin B/pharmacology
15.
Braz. j. microbiol ; 48(3): 493-498, July-Sept. 2017. tab, graf
Article in English | LILACS | ID: biblio-889153

ABSTRACT

Abstract Carbapenems are considered last-line agents for the treatment of serious infections caused by Klebsiella pneumoniae, and this microorganism may exhibit resistance to β-lactam antibiotics due to different mechanisms of resistance. We evaluated 27 isolates of K. pneumoniae resistant to carbapenems recovered from inpatients at the University Hospital of Santa Maria-RS from July 2013 to August 2014. We carried out antimicrobial susceptibility, carbapenemase detection, testing for the presence of efflux pump by broth microdilution and loss of porin by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Genetic similarity was evaluated by ERIC-PCR. High levels of resistance were verified by the minimum inhibitory concentration for the antimicrobials tested. The blaKPC gene was present in 89% of the clinical isolates. Blue-Carba and combined disk with AFB tests showed 100% concordance, while the combined disk test with EDTA showed a high number of false-positives (48%) compared with the gold-standard genotypic test. Four isolates showed a phenotypic resistance profile consistent with the overexpression of the efflux pump, and all clinical isolates had lost one or both porins. The ERIC-PCR dendrogram demonstrated the presence of nine clusters. The main mechanism of resistance to carbapenems found in the assessed isolates was the presence of the blaKPC gene.


Subject(s)
Humans , Anti-Bacterial Agents/pharmacology , Carbapenems/pharmacology , Klebsiella Infections/microbiology , Klebsiella pneumoniae/drug effects , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Drug Resistance, Bacterial , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/isolation & purification , Klebsiella pneumoniae/metabolism , Microbial Sensitivity Tests
16.
Rev. Soc. Bras. Med. Trop ; 50(3): 321-328, May-June 2017. tab, graf
Article in English | LILACS | ID: biblio-896978

ABSTRACT

Abstract INTRODUCTION: Infections caused by β-lactamase-producing gram-negative bacteria, such as Klebsiella pneumoniae, are increasing globally with high morbidity and mortality. The aim of the current study was to determine antimicrobial susceptibility patterns and the prevalence of antibiotic resistance genes (β-lactamase and integron genes) using multiplex PCR. METHODS One-hundred K. pneumoniae isolates were collected from different clinical samples. Antibiotic susceptibility testing was performed with thirteen different antibiotics. Multiplex-PCR was used to detect β-lactamase (bla TEM, bla CTX-M, bla SHV , bla VEB, bla PER, bla GES, bla VIM, bla IMP, bla OXA, and bla KPC) and integron genes (int I, int II, and int III). RESULTS: The highest and lowest rate of resistance was exhibited against amikacin (93%) and imipenem (8%), respectively. The frequency of β-lactamase-positive K. pneumoniae was 37%, and the prevalence of the bla TEM, bla CTX-M, bla SHV , bla VEB, bla PER, bla GES, bla VIM, bla IMP, bla OXA, and bla KPC genes was 38%, 24%, 19%, 12%, 6%, 11%, 33%, 0%, 28%, and 23%, respectively. Of the 100 isolates, eight (8%) were positive for class I integrons; however, class II and III integrons were not detected in any of the strains. CONCLUSIONS: These results indicate co-carriage of a number of β-lactamase genes and antibiotic resistance integrons on the same plasmids harboring multi-drug resistance genes. It seems that these properties help to decrease treatment complications due to resistant bacterial infections by rapid detection, infection-control programs and prevention of transmission of drug resistance.


Subject(s)
Humans , beta-Lactamases/genetics , Drug Resistance, Bacterial/genetics , Integrons/genetics , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/genetics , Klebsiella Infections/microbiology , DNA, Bacterial/genetics , Microbial Sensitivity Tests , Cross-Sectional Studies , Multiplex Polymerase Chain Reaction , Klebsiella pneumoniae/isolation & purification , Anti-Bacterial Agents/pharmacology
17.
Rev. chil. infectol ; 33(5): 519-523, oct. 2016. ilus, graf, tab
Article in Spanish | LILACS | ID: biblio-844403

ABSTRACT

In order to study the clonal relationship and blaKPC gene detection in clinical isolates of Klebsiella pneumoniae resistant to carbapenems, we analyzed 22 clinical isolates of K. pneumoniae with resistance to imipenem and/ or meropenem, isolated in the laboratory of bacteriology at the University Hospital "Antonio Patricio de Alcalá" (HUAPA) from the Cumana city, Sucre state, Venezuela, for a period of five consecutive years. Susceptibility to different antimicrobials was determined, and the presence of carbapenemases was detected by modified Hodge method, phenyl boronic acid synergy and combination discs. blaKPC gene detection was conducted by polymerase chain reaction and the clonal relationship was determined by pulsed field electrophoresis. High rates of antimicrobial resistance were found, five strains were negative, at least one phenotypic method, and all carried the blaKPC gene. Clonal spread was observed only in the intensive care unit (ICU), while in other services, polyclonality was found. We concluded that blaKPC gene is present in K. pneumoniae strains resistant to carbapenems isolated in the HUAPA and clonal spread it was only in the ICU.


Con el objetivo de estudiar la relación clonal y detección del gen blaKPC en aislados clínicos de Klebsiella pneumoniae resistentes a carbapenémicos, se analizaron 22 cepas clínicas de K. pneumoniae con resistencia a imipenem y/o meropenem, aisladas en el laboratorio de bacteriología del Hospital Universitario "Antonio patricio de Alcalá" (HUAPA) de la ciudad de cumaná, Estado Sucre, Venezuela, durante un período de cinco años continuos. Se determinó la susceptibilidad a diversos antimicrobianos, y se detectó la presencia de carbapenemasas por los métodos de Hodge modificado, sinergia con ácido fenil borónico y combinación de discos. La detección del gen blaKPC se llevó a cabo mediante la técnica de reacción de polimerasa en cadena y la determinación de la relación clonal se realizó por electroforesis de campo pulsado. Se encontraron elevados porcentajes de resistencia antimicrobiana, cinco cepas resultaron negativas, al menos, a un método fenotípico y todas portaban el gen blaKPC. Se observó diseminación de clones únicamente en la Unidad de cuidados Intensivos (UCI), mientras que, en otros servicios, se halló policlonalidad. Se concluye que el gen blaKPC se encuentra presente en cepas de K. pneumoniae resistentes a carbapenémicos aisladas en el HUAPA y que hubo diseminación clonal sólo en UCI.


Subject(s)
Humans , beta-Lactamases/genetics , Klebsiella Infections/microbiology , Carbapenems/pharmacology , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/genetics , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Venezuela , DNA, Bacterial/genetics , Microbial Sensitivity Tests , Polymerase Chain Reaction , Electrophoresis, Gel, Pulsed-Field , Clone Cells
18.
Rev. Soc. Bras. Med. Trop ; 49(4): 441-445, July-Aug. 2016. tab, graf
Article in English | LILACS | ID: lil-792801

ABSTRACT

Abstract: INTRODUCTION: Extended-spectrum beta-lactamases (ESBLs) are bacterial enzymes capable of hydrolyzing beta-lactams. The aim of this study was to describe the prevalence of TEM- and SHV-type ESBL-producing Klebsiella pneumoniae strains in Zahedan, Southeast Iran. METHODS: A total of 170 non-repetitive K. pneumoniae strains were collected from patients referred to three teaching hospitals of Zahedan. Antibiotic susceptibility testing was determined for 17 antibiotics using the Kirby-Bauer disc diffusion method. The frequency of ESBL-producing strains was calculated, and minimum inhibitory concentrations of ESBL-producing strains were determined for cefotaxime, ceftazidime, ceftriaxone, and cefpodoxime. The presence of bla TEM and bla SHV genes was tested in all ESBL-producing strains using polymerase chain reaction and DNA sequencing. RESULTS: Among the 170 K. pneumoniae clinical isolates, 55 (32.4%) were ESBL producers; 92.7% (n=51) and 72.7% (n=40) of the isolates carried the bla SHV and bla TEM genes, respectively, and 67.3% (n=37) carried both genes. The sequencing results showed that all bla TEM types were bla TEM-1, except for two isolates that were bla TEM-104. The bla SHV types were bla SHV-1, bla SHV-11, bla SHV-12, bla SHV-99, bla SHV-108, and bla SHV-110. CONCLUSIONS: The percentage of bla TEM and bla SHV among ESBL-producing K. pneumoniae isolates from Zahedan is relatively high, indicating the need for further surveillance and consideration in antibiotic use. To the best of our knowledge, this is the first report of TEM-104-, SHV-99-, SHV-108-, and SHV-110-type ESBLs among clinical isolates of K. pneumoniae from Iran, and TEM-1, SHV-1, SHV-11, and SHV-12 appear to be the dominant ESBLs in this region.


Subject(s)
Humans , Bacterial Proteins/genetics , beta-Lactamases/biosynthesis , DNA, Bacterial/genetics , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/genetics , Anti-Bacterial Agents/pharmacology , Phenotype , Klebsiella Infections/microbiology , Sequence Analysis, DNA , Drug Resistance, Multiple, Bacterial/genetics , Disk Diffusion Antimicrobial Tests , Genes, Bacterial , Genotype , Iran
19.
Braz. j. infect. dis ; 20(2): 134-140, Mar.-Apr. 2016. tab, graf
Article in English | LILACS | ID: lil-780802

ABSTRACT

Abstract Background Little is known about factors associated with carbapenem-resistant Klebsiella pneumoniae infections in pediatric patients, who are initally colonized with carbapenem-resistant Klebsiella pneumoniae. Materials and methods A retrospective case–control study was conducted involving pediatric and neonatal intensive care units throughout a five-year period (January 2010–December 2014). Clinical and microbiological data were extracted from Hospital Infection Control Committee reports and patients’ medical records. Risk factors were assessed in carbapenem-resistant Klebsiella pneumoniae colonized patients who developed subsequent systemic infection (cases) and compared to carbapenem-resistant Klebsiella pneumoniae colonized patients who did not develop infection (controls). Results Throughout the study period, 2.6% of patients admitted to neonatal intensive care units and 3.6% of patients admitted to pediatric intensive care units had become colonized with carbapenem-resistant Klebsiella pneumoniae. After a mean of 10.6 ± 1.9 days (median: 7 days, range: 2–38 days) following detection of colonization, 39.0% of the carbapenem-resistant Klebsiella pneumoniae colonized patients in pediatric intensive care units and 18.1% of carbapenem-resistant Klebsiella pneumoniae colonized patients in neonatal intensive care units developed systemic carbapenem-resistant Klebsiella pneumoniae infection. Types of systemic carbapenem-resistant Klebsiella pneumoniae infections included bacteremia (n = 15, 62.5%), ventilator-associated pneumonia (n = 4, 16.6%), ventriculitis (n = 2, 8.3%), intraabdominal infections (n = 2, 8.3%), and urinary tract infection (n = 1, 4.1%). A logistic regression model including parameters found significant in univariate analysis of carbapenem resistant Klebsiella pneumoniae colonization and carbapenem resistant Klebsiella pneumoniae infection groups revealed underlying metabolic disease (OR: 10.1; 95% CI: 2.7–37.2), previous carbapenem use (OR: 10.1; 95% CI: 2.2–40.1), neutropenia (OR: 13.8; 95% CI: 3.1–61.0) and previous surgical procedure (OR: 7.4; 95% CI: 1.9–28.5) as independent risk factors for carbapenem-resistant Klebsiella pneumoniae infection in patients colonized with carbapenem-resistant Klebsiella pneumoniae. Out of 24 patients with carbapenem resistant Klebsiella pneumoniae infection, 4 (16.6%) died of carbapenem-resistant Klebsiella pneumoniae sepsis. Conclusion Asymptomatic colonization with carbapenem-resistant Klebsiella pneumoniae in intensive care units of pediatric departments should alert health care providers about forthcoming carbapenem-resistant Klebsiella pneumoniae infection. Those carbapenem-resistant Klebsiella pneumoniae colonized patients at risk of developing infection due to carbapenem-resistant Klebsiella pneumoniae may be targeted for interventions to reduce subsequent infection occurence and also for timely initiation of empirical carbapenem-resistant Klebsiella pneumoniae active treatment, when necessary.


Subject(s)
Humans , Male , Female , Infant, Newborn , Infant , Child, Preschool , Child , Adolescent , Klebsiella Infections/microbiology , Carbapenems/pharmacology , Cross Infection/microbiology , Drug Resistance, Bacterial , Klebsiella pneumoniae/drug effects , Anti-Bacterial Agents/pharmacology , Rectum/microbiology , Klebsiella Infections/epidemiology , Intensive Care Units, Neonatal , Cross Infection/epidemiology , Epidemiologic Methods , Disease Progression , beta-Lactam Resistance , Disk Diffusion Antimicrobial Tests , Klebsiella pneumoniae/isolation & purification
20.
Rev. Soc. Bras. Med. Trop ; 49(2): 177-182, Mar.-Apr. 2016. tab, graf
Article in English | LILACS | ID: lil-782100

ABSTRACT

Abstract: INTRODUCTION Klebsiella pneumoniae has become an increasingly important etiologic agent of nosocomial infections in recent years. This is mainly due to the expression of virulence factors and development of resistance to several antimicrobial drugs. METHODS This retrospective study examines data obtained from the microbiology laboratory of a Brazilian tertiary-care hospital. To assess temporal trends in prevalence and antimicrobial susceptibility, K. pneumoniae isolates were analyzed from 2000 to 2013. The relative frequencies of K. pneumoniae isolation were calculated among all Gram-negative bacilli isolated in each period analyzed. Susceptibility tests were performed using automated systems. RESULTS: From 2000-2006, K. pneumonia isolates comprised 10.7% of isolated Gram-negative bacilli (455/4260). From 2007-2013, this percentage was 18.1% (965/5331). Strictly considering isolates from bloodstream infections, the relative annual prevalence of K. pneumoniae increased from 14-17% to 27-32% during the same periods. A progressive decrease in K. pneumoniae susceptibility to all antimicrobial agents assessed was detected. Partial resistance was also observed to antimicrobial drugs that have been used more recently, such as colistin and tigecycline. CONCLUSIONS Our study indicates that K. pneumoniae has become a major pathogen among hospitalized patients and confirms its recent trend of increasing antimicrobial resistance.


Subject(s)
Humans , Klebsiella Infections/microbiology , Klebsiella pneumoniae/drug effects , Anti-Bacterial Agents/pharmacology , Brazil , Microbial Sensitivity Tests , Prevalence , Retrospective Studies , Drug Resistance, Bacterial/drug effects , Tertiary Care Centers/statistics & numerical data , Klebsiella pneumoniae/isolation & purification
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