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The dissemination of blaKPC-harboring Pseudomonas aeruginosa (KPC-Pa) is considered a serious public health problem. This study provides an overview of the epidemiology of these isolates to try to elucidate novel mobilization platforms that could contribute to their worldwide spread. A systematic review in PubMed and EMBASE was performed to find articles published up to June 2022. In addition, a search algorithm using NCBI databases was developed to identify sequences that contain possible mobilization platforms. After that, the sequences were filtered and pair-aligned to describe the blaKPC genetic environment. We found 691 KPC-Pa isolates belonging to 41 different sequence types and recovered from 14 countries. Although the blaKPC gene is still mobilized by the transposon Tn4401, the non-Tn4401 elements (NTEKPC) were the most frequent. Our analysis allowed us to identify 25 different NTEKPC, mainly belonging to the NTEKPC-I, and a new type (proposed as IVa) was also observed. This is the first systematic review that consolidates information about the behavior of the blaKPC acquisition in P. aeruginosa and the genetic platforms implied in its successful worldwide spread. Our results show high NTEKPC prevalence in P. aeruginosa and an accelerated dynamic of unrelated clones. All information collected in this review was used to build an interactive online map.
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Most current anti-viral vaccines elicit a humoral and cellular immune response via the pathway of phagocytic cell mediated viral antigen presentation to B and T cell surface receptors. However, this pathway results in reduced ability to neutralize S-protein Receptor Binding Domains (RBDs) from several Variants of Concern (VOC) and the rapid waning of memory B cell response requiring vaccine reformulation to cover dominant VOC S-proteins and multiple boosters. Here we show for the first time in mice and humans, that a bacterially derived, non-living, nanocell (EDV; EnGeneIC Dream Vector) packaged with plasmid expressed SARS-CoV-2 S-protein and α-galactosyl ceramide adjuvant (EDV-COVID-αGC), stimulates an alternate pathway due to dendritic cells (DC) displaying both S-polypeptides and αGC thereby recruiting and activating iNKT cells with release of IFNγ. This triggers DC activation/maturation, activation of follicular helper T cells (TFH), cognate help to B cells with secretion of a cytokine milieu promoting B cell maturation, somatic hypermutation in germinal centers to result in high affinity antibodies. Surrogate virus neutralization tests show 90-100% neutralization of ancestral and early VOC in mice and human trial volunteers. EDV-COVID-αGC as a third dose booster neutralized Omicron BA. 4/5. Serum and PBMC analyses reveal long lasting S-specific memory B and T cells. In contrast, control EDVs lacking αGC, did not engage the iNKT/DC pathway resulting in antibody responses unable to neutralize all VOCs and had a reduced B cell memory. The vaccine is lyophilized, stored and transported at room temperature with a shelf-life of over a year.
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Vacinas contra COVID-19 , COVID-19 , Humanos , Animais , Camundongos , Leucócitos Mononucleares , SARS-CoV-2 , Apresentação de AntígenoRESUMO
Resistance to carbapenems in Klebsiella pneumoniae has been mostly related with the worldwide dissemination of KPC, largely due to the pandemic clones belonging to the complex clonal (CC) 258. To unravel blaKPC post-endemic clinical impact, here we describe clinical characteristics of 68 patients from a high complexity hospital, and the molecular and genetic characteristics of their 139 blaKPC-K. pneumoniae (KPC-Kp) isolates. Of the 26 patients that presented relapses or reinfections, 16 had changes in the resistance profiles of the isolates recovered from the recurrent episodes. In respect to the genetic diversity of KPC-Kp isolates, PFGE revealed 45 different clonal complexes (CC). MLST for 12 representative clones showed ST258 was present in the most frequent CC (23.0%), however, remaining 11 representative clones belonged to non-CC258 STs (77.0%). Interestingly, 16 patients presented within-patient genetic diversity of KPC-Kp clones. In one of these, three unrelated KPC-Kp clones (ST258, ST504, and ST846) and a blaKPC-K. variicola isolate (ST182) were identified. For this patient, complete genome sequence of one representative isolate of each clone was determined. In K. pneumoniae isolates blaKPC was mobilized by two Tn3-like unrelated platforms: Tn4401b (ST258) and Tn6454 (ST504 and ST846), a new NTEKPC-IIe transposon for first time characterized also determined in the K. variicola isolate of this study. Genome analysis showed these transposons were harbored in different unrelated but previously reported plasmids and in the chromosome of a K. pneumoniae (for Tn4401b). In conclusion, in the blaKPC post-endemic dissemination in Colombia, different KPC-Kp clones (mostly non-CC258) have emerged due to integration of the single blaKPC gene in new genetic platforms. This work also shows the intra-patient resistant and genetic diversity of KPC-Kp isolates. This circulation dynamic could impact the effectiveness of long-term treatments.
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Proteínas de Bactérias/genética , Carbapenêmicos/farmacologia , Farmacorresistência Bacteriana , Klebsiella pneumoniae/genética , Tipagem de Sequências Multilocus/instrumentação , beta-Lactamases/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Colômbia , Feminino , Variação Genética , Genoma Bacteriano , Genômica , Hospitalização , Hospitais , Humanos , Infecções por Klebsiella , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Estudos Retrospectivos , Sequenciamento Completo do Genoma , Adulto JovemRESUMO
Immunotherapy has emerged as a powerful new chapter in the fight against cancer. However, it has yet to reach its full potential due in part to the complexity of the cancer immune response. We demonstrate that tumor-targeting EDV nanocells function as an immunotherapeutic by delivering a cytotoxin in conjunction with activation of the immune system. These nanocells polarize M1 macrophages and activate NK cells concurrently producing a Th1 cytokine response resulting in potent antitumor function. Dendritic cell maturation and antigen presentation follows, which generates tumor-specific CD8+ T cells, conferring prolonged tumor remission. The combination of cytotoxin delivery and activation of innate and adaptive antitumor immune responses results in a potent cyto-immunotherapeutic with potential in clinical oncology.
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Antineoplásicos/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Imunidade Inata/efeitos dos fármacos , Salmonella typhimurium/citologia , Adulto , Idoso , Animais , Antineoplásicos/uso terapêutico , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/patologia , Carcinoma Ductal Pancreático/tratamento farmacológico , Linhagem Celular , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/fisiologia , Doxorrubicina/administração & dosagem , Doxorrubicina/análogos & derivados , Receptores ErbB/administração & dosagem , Receptores ErbB/metabolismo , Feminino , Glioblastoma/tratamento farmacológico , Glioblastoma/patologia , Humanos , Imunoterapia/métodos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Nanoestruturas/química , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/patologiaRESUMO
Human malignant mesothelioma is a chemoresistant tumour that develops from mesothelial cells, commonly associated with asbestos exposure. Malignant mesothelioma incidence rates in European countries are still rising and Australia has one of the highest burdens of malignant mesothelioma on a population basis in the world. Therapy using systemic delivery of free cytotoxic agents is associated with many undesirable side effects due to non-selectivity, and is thus dose-limited which limits its therapeutic potential. Therefore, increasing the selectivity of anti-cancer agents has the potential to dramatically enhance drug efficacy and reduce toxicity. EnGeneIC Dream Vectors (EDV) are antibody-targeted nanocells which can be loaded with cytotoxic drugs and delivered to specific cancer cells via bispecific antibodies (BsAbs) which target the EDV and a cancer cell-specific receptor, simultaneously. BsAbs were designed to target doxorubicin-loaded EDVs to cancer cells via cell surface mesothelin (MSLN). Flow cytometry was used to investigate cell binding and induction of apoptosis, and confocal microscopy to visualize internalization. Mouse xenograft models were used to assess anti-tumour effects in vivo, followed by immunohistochemistry for ex vivo evaluation of proliferation and necrosis. BsAb-targeted, doxorubicin-loaded EDVs were able to bind to and internalize within mesothelioma cells in vitro via MSLN receptors and induce apoptosis. In mice xenografts, the BsAb-targeted, doxorubicin-loaded EDVs suppressed the tumour growth and also decreased cell proliferation. Thus, the use of MSLN-specific antibodies to deliver encapsulated doxorubicin can provide a novel and alternative modality for treatment of mesothelioma.
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Proliferação de Células , Mesotelioma/patologia , Receptores de Superfície Celular/metabolismo , Animais , Humanos , Mesotelina , Camundongos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Bacteria that produce the broad-spectrum Carbapenem antibiotic New Delhi Metallo-ß-lactamase (NDM) place a burden on health care systems worldwide, due to the limited treatment options for infections caused by them and the rapid global spread of this antibiotic resistance mechanism. Although it is believed that the associated resistance gene blaNDM-1 originated in Acinetobacter spp., the role of Enterobacteriaceae in its dissemination remains unclear. In this study, we used whole genome sequencing to investigate the dissemination dynamics of blaNDM-1-positive plasmids in a set of 21 clinical NDM-1-positive isolates from Colombia and Mexico (Providencia rettgeri, Klebsiella pneumoniae, and Acinetobacter baumannii) as well as six representative NDM-1-positive Escherichia coli transconjugants. Additionally, the plasmids from three representative P. rettgeri isolates were sequenced by PacBio sequencing and finished. Our results demonstrate the presence of previously reported plasmids from K. pneumoniae and A. baumannii in different genetic backgrounds and geographically distant locations in Colombia. Three new previously unclassified plasmids were also identified in P. rettgeri from Colombia and Mexico, plus an interesting genetic link between NDM-1-positive P. rettgeri from distant geographic locations (Canada, Mexico, Colombia, and Israel) without any reported epidemiological links was discovered. Finally, we detected a relationship between plasmids present in P. rettgeri and plasmids from A. baumannii and K. pneumoniae. Overall, our findings suggest a Russian doll model for the dissemination of blaNDM-1 in Latin America, with P. rettgeri playing a central role in this process, and reveal new insights into the evolution and dissemination of plasmids carrying such antibiotic resistance genes.
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Infecções por Acinetobacter/microbiologia , Acinetobacter baumannii/enzimologia , Proteínas de Bactérias/genética , Infecções por Enterobacteriaceae/microbiologia , Enterobacteriaceae/enzimologia , Plasmídeos/genética , beta-Lactamases/genética , Infecções por Acinetobacter/epidemiologia , Acinetobacter baumannii/classificação , Acinetobacter baumannii/efeitos dos fármacos , Acinetobacter baumannii/genética , Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Colômbia/epidemiologia , Farmacorresistência Bacteriana , Enterobacteriaceae/classificação , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/genética , Infecções por Enterobacteriaceae/epidemiologia , Humanos , México/epidemiologia , Filogenia , Plasmídeos/metabolismo , beta-Lactamases/metabolismoRESUMO
BACKGROUND: Community-genotype methicillin-resistant Staphylococcus aureus (CG-MRSA) clones are a global concern due to their resistance and increased virulence and their ability to cause infections both hospitalized patients and healthy people in the community. Here, we characterize 32 isolates of a new CG-MRSA clone. These isolates were identified in four cities in Colombia, South America. METHODS: The isolates were recovered from four different epidemiological and prospective studies that were conducted in several regions of Colombia. Molecular characterizations included multilocus sequence typing; pulsed-field gel electrophoresis; SCCmec, agr and spa typing; and whole-genome sequencing. RESULTS: All isolates belonged to ST923 (clonal complex 8), harbouring SCCmec IVa and a spa type t1635 and lacking an arginine catabolism mobile element. The isolates were classified as COL923, were resistant to at least one non-beta-lactam antibiotic, and exhibited high frequencies (>60%) of resistance to macrolides and tetracycline. Using whole-genome sequencing, we found that this new clone harbours novel prophage 3 and beta-island structures and a slightly different pathogenicity island 5. Moreover, isolates belonging to the COL923 clone are grouped in a different clade than USA300 and USA300-LV. CONCLUSION: Our results show the emergence and spread of the COL923 clone in different cities in Colombia. This clone is resistant to several antibiotics and possesses new structures in its mobile genetic elements.
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Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Infecções Estafilocócicas/epidemiologia , Adolescente , Antibacterianos , Criança , Pré-Escolar , Colômbia/epidemiologia , Feminino , Genótipo , Humanos , Lactente , Recém-Nascido , Masculino , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/patogenicidade , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Estudos Prospectivos , Infecções Estafilocócicas/microbiologia , Virulência/genéticaRESUMO
Providencia rettgeri is an opportunistic bacterial pathogen of clinical significance due to its association with urinary tract infections and multidrug resistance. Here, we report the first complete genome sequence of P. rettgeri The genome of strain RB151 consists of a 4.8-Mbp chromosome and a 108-kbp blaNDM-1-positive plasmid.
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Drug resistant Pseudomonas aeruginosa represents a therapeutic challenge. To assess the diversity of P. aeruginosa antibiotic resistant variants, isolates were recovered from hospital patients in Colombia. Thirty of 60 isolates contained class 1 integrons and five were of Sequence Type ST235 having appeared in a single intensive care unit. All five possessed an unusual integron but showed differences in gene cassette content and the presence/absence of insertion sequence IS26. This showed that differences can arise rapidly, even within a single ICU. Also, the emergence of IS26 in P. aeruginosa is contributing to the evolution of resistance in this bacterium.
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Infecção Hospitalar/microbiologia , Farmacorresistência Bacteriana Múltipla , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/isolamento & purificação , Antibacterianos/farmacologia , Colômbia/epidemiologia , Infecção Hospitalar/epidemiologia , Humanos , Unidades de Terapia Intensiva/estatística & dados numéricos , Testes de Sensibilidade Microbiana , Infecções por Pseudomonas/epidemiologia , Pseudomonas aeruginosa/classificação , Pseudomonas aeruginosa/genéticaRESUMO
OBJECTIVE: Community-genotype methicillin-resistant Staphylococcus aureus (CG-MRSA) isolates are known to be more virulent and clinically aggressive in children. The goal of the present study was characterize the molecular epidemiology of MRSA isolates causing infections in Colombian children. METHODS: An observational and prospective study was conducted between April 2009 and June 2011 at 15 hospitals in Bogotá, Colombia. A detailed epidemiological profile was made of 162 children infected with MRSA. The isolates were subjected to antimicrobial susceptibility testing, molecular characterization including 21 virulence genes, SCCmec, spa and agr typing, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). RESULTS: Among all isolates included in the study, 85.8% were obtained from patients whose infectious process was initiated in the community; of these, 69,8% occurred in patients without healthcare-associated risk factors. The molecular characterization of the isolates showed a high proportion (95.1%) containing a community-genotype profile with a high prevalence of SCCmec type IV, PVL-positives, and also related to CC8. Most CG-MRSA isolates (143, 92.9%) were genetically related to the pandemic clone USA300, differing by the presence of SCCmec IVc and the absence of the arginine catabolic mobile element (ACME). CONCLUSIONS: An increase in the frequency of CG-MRSA infections has been reported worldwide. In this study we found that almost all MRSA infections in our pediatric population were caused by community-genotype isolates, supporting the success of the CG-MRSA clones.
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Infecções Comunitárias Adquiridas , Infecção Hospitalar , Staphylococcus aureus Resistente à Meticilina/genética , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Antibacterianos/farmacologia , Criança , Pré-Escolar , Colômbia/epidemiologia , Eletroforese em Gel de Campo Pulsado , Humanos , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Estudos ProspectivosRESUMO
INTRODUCTION: USA300 is a genetic lineage found both in methicillin-resistant (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) isolates. In Colombia, hospital and community MRSA infections are caused by a USA300-related community genotype MRSA (CG-MRSA) clone. The genetic origin of this clone is unknown yet. OBJECTIVE: To identify and characterize methicillin-resistant (MRSA) and methicillin-sensitive S. aureus (MSSA) isolates in order to improve the information about the origin of the CG-MRSA isolates in Colombia. MATERIALS AND METHODS: USA300-related MSSA isolates were detected and characterized from a study of 184 S. aureus isolates (90 MRSA and 94 MSSA) recovered from infections. The genetic relatedness of the isolates was established by means of pulsed field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and protein A gene typification ( spa typing). RESULTS: Among 184 isolates, 27 (14.7%) showed molecular characteristics and genetic relationship with the USA300 clone, of which 18 were MRSA and nine were MSSA. All USA300-related MRSA harbored Staphylococcal cassette chromosome mec (SCC mec ) IVc (3.1.2). In the MSSA isolates, SCC mec remnants or att B duplicate sites were not detected. CONCLUSIONS: In Colombia, the CG-MRSA isolates probably originated in the dissemination of an USA300-related MSSA clone which later acquired SCC mec IVc.
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Infecções Comunitárias Adquiridas/microbiologia , Resistência a Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/genética , Infecções Estafilocócicas/microbiologia , Adolescente , Adulto , Idoso , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Chile , Células Clonais , Colômbia/epidemiologia , Infecções Comunitárias Adquiridas/epidemiologia , Infecções Comunitárias Adquiridas/transmissão , Farmacorresistência Bacteriana Múltipla/genética , Eletroforese em Gel de Campo Pulsado , Genes Bacterianos , Genótipo , Humanos , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/patogenicidade , Pessoa de Meia-Idade , Proteínas de Ligação às Penicilinas , Filogenia , Estudos Prospectivos , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/transmissão , Estados Unidos , Virulência/genética , Adulto JovemRESUMO
Introducción. USA300 es un linaje genético que se encuentra en aislamientos de Staphylococcus aureus sensibles (SASM) y resistentes a meticilina (SARM). Actualmente, en Colombia las infecciones por SARM en hospitales y en la comunidad son causadas principalmente por un clon con genotipo comunitario (SARM-GC) relacionado genéticamente con el clon USA300. El origen de esta variante es aún desconocido. Objetivo. Identificar y caracterizar aislamientos de S. aureus resistentes y sensibles a meticilina con el fin de aportar información para establecer un posible origen de los aislamientos SARM-GC en Colombia. Materiales y métodos. Se realizó una caracterización de aislamientos SASM relacionados con el clon USA300 detectados a partir de un análisis de 184 aislamientos de S. aureus (90 SARM y 94 SASM) causantes de infecciones. La relación genética de los aislamientos se determinó por electroforesis en gel de campo pulsado (PFGE), tipificación de secuencias multilocus (MLST) y tipificación del gen de la proteína A ( spa ). Resultados. De los 184 aislamientos, 27 (14,7 %) presentaron características moleculares y relación genética con el clon USA300, y de ellos, 18 fueron SARM y nueve fueron SASM. Todos los aislamientos SARM relacionados con este clon albergaban un casete estafilocócico cromosómico mec (SCC mec ) IVc (3.1.2). En ningún aislamiento SASM se detectaron secuencias remanentes de SCC mec o una duplicación del sitio att B que evidenciaran la pérdida del casete. Conclusión. El origen de los aislamientos SARM-GC en Colombia probablemente se encuentre en la diseminación de clones SASM relacionados con el clon USA300 que adquirieron el SCC mec IVc posteriormente.
Introduction: USA300 is a genetic lineage found both in methicillin-resistant (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) isolates. In Colombia, hospital and community MRSA infections are caused by a USA300-related community genotype MRSA (CG-MRSA) clone. The genetic origin of this clone is unknown yet. Objective: To identify and characterize methicillin-resistant (MRSA) and methicillin-sensitive S. aureus (MSSA) isolates in order to improve the information about the origin of the CG-MRSA isolates in Colombia. Materials and methods: USA300-related MSSA isolates were detected and characterized from a study of 184 S. aureus isolates (90 MRSA and 94 MSSA) recovered from infections. The genetic relatedness of the isolates was established by means of pulsed field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and protein A gene typification ( spa typing). Results: Among 184 isolates, 27 (14.7%) showed molecular characteristics and genetic relationship with the USA300 clone, of which 18 were MRSA and nine were MSSA. All USA300-related MRSA harbored Staphylococcal cassette chromosome mec (SCC mec ) IVc (3.1.2). In the MSSA isolates, SCC mec remnants or att B duplicate sites were not detected. Conclusions: In Colombia, the CG-MRSA isolates probably originated in the dissemination of an USA300-related MSSA clone which later acquired SCC mec IVc.
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Adolescente , Adulto , Idoso , Humanos , Pessoa de Meia-Idade , Adulto Jovem , Infecções Comunitárias Adquiridas/microbiologia , Resistência a Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/genética , Infecções Estafilocócicas/microbiologia , Técnicas de Tipagem Bacteriana , Proteínas de Bactérias/genética , Chile , Células Clonais , Colômbia/epidemiologia , Infecções Comunitárias Adquiridas/epidemiologia , Infecções Comunitárias Adquiridas/transmissão , Farmacorresistência Bacteriana Múltipla/genética , Eletroforese em Gel de Campo Pulsado , Genes Bacterianos , Genótipo , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/patogenicidade , Filogenia , Estudos Prospectivos , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/transmissão , Estados Unidos , Virulência/genéticaRESUMO
ß-Lactam resistance in Pseudomonas aeruginosa clinical isolates is driven by a number of mechanisms. Whilst several are understood, how they act co-operatively in pathogenic strains is less clear. In some isolates, resistance profiles cannot always be explained by identifying the common resistance-determining pathways, suggesting that other mechanisms may be important. Pathogenic P. aeruginosa isolates from four countries were characterised by PCR. Quantitative expression analysis was also assessed for the activity of several pathways that influence antibiotic resistance, and culture experiments were conducted to test how random transposition of the insertion sequence IS26 during growth may influence resistance to some antibiotics. In most strains, antibiotic resistance was being driven by changes in multiple pathways and by the presence or absence of genes acquired by lateral gene transfer. Multiple mechanisms of resistance were prevalent in strains from all of the countries examined, although regional differences in the type of interacting mechanisms were apparent. Changes in chromosomal pathways included overexpression of AmpC and two efflux pumps. Also, gain or loss of IS26 at some chromosomal locations, most notably oprD, could influence resistance to carbapenems. IS26-related resistance was found in strains from Argentina and geographically linked Uruguay, but not in strains from either Colombia or Australia. Pseudomonas aeruginosa pathogenic strains are evolving to become multidrug-resistant in more complex ways. This is being influenced by single strains acquiring changes in numerous known pathways as well as by newly emerging resistance mechanisms in this species.
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The dissemination of a clone of community genotype methicillin-resistant Staphylococcus aureus (CG-MRSA) that is related to USA300 has been reported in Latin America. We recently detected isolates of a new clone of CG-MRSA (spa type t1635 and ACME-negative) that was genetically unrelated to the USA300 clone and that causes infections in children in Colombia. This finding indicates the appearance of a new clone of CG-MRSA in our region.
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Genótipo , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/genética , Infecções Estafilocócicas/diagnóstico , Adolescente , Criança , Pré-Escolar , Colômbia/epidemiologia , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Infecções Estafilocócicas/epidemiologiaRESUMO
INTRODUCTION: Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) infections are found with increasing the frequency, both in healthy individuals in the community and in hospitalized patients. In Colombia and the Andean region, CA-MRSA isolates have a genetic background that is related to the pandemic USA300 clone. OBJECTIVE: Two molecular methods are designed and standardized for the rapid differentiation of Colombian community-acquired and hospital-acquired methicillin-resistant Staphylococcus aureus (HA-MRSA) isolates. MATERIALS AND METHODS: Two molecular methods were standardized for the identification of CA-MRSA isolates. The first method was based on the differential digestion of the carbamate kinase (arcC)and guanylate kinase (gmk) genes in the sequences type 5 (ST5) in the HA-MRSA isolates and 8 (ST8) in the CA-MRSA isolates. The second method was based on the PCR amplification of 5 specific virulence factors found in CA-MRSA and HA-MRSA isolates. The specificity and precision of each method were evaluated using 237 clinical MRSA isolates. RESULTS: The first method identified 100% and 93.2% of the CA-MRSA and HA-MRSA isolates, respectively. The second method also correctly identified the two isolates types (CA-MRSA and HA-MRSA). CONCLUSIONS: These two methods are a convenient alternative for the rapid identification of the CA-MRSA isolates, compared with other techniques such as pulsed field gel electrophoresis and multilocus sequence typing, which are time-consuming and more expensive.
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Técnicas de Tipagem Bacteriana/métodos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de Restrição , Infecções Estafilocócicas/microbiologia , Alelos , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana/normas , Colômbia , Infecções Comunitárias Adquiridas/epidemiologia , Infecções Comunitárias Adquiridas/microbiologia , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Eletroforese em Gel de Campo Pulsado , Genes Bacterianos , Guanilato Quinases/genética , Humanos , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/genética , Fosfotransferases (Aceptor do Grupo Carboxila)/genética , Reprodutibilidade dos Testes , Alinhamento de Sequência , Análise de Sequência de DNA , Infecções Estafilocócicas/epidemiologia , Fatores de Tempo , Virulência/genéticaRESUMO
Introduction. Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) infections are found with increasing the frequency, both in healthy individuals in the community and in hospitalized patients. In Colombia and the Andean region, CA-MRSA isolates have a genetic background that is related to the pandemic USA300 clone. Objective. Two molecular methods are designed and standardized for the rapid differentiation of Colombian community-acquired and hospital-acquired methicillin-resistant Staphylococcus aureus (HA-MRSA) isolates. Materials and methods. Two molecular methods were standardized for the identification of CA-MRSA isolates. The first method was based on the differential digestion of the carbamate kinase (arcC)and guanylate kinase (gmk) genes in the sequences type 5 (ST5) in the HA-MRSA isolates and 8 (ST8) in the CA-MRSA isolates. The second method was based on the PCR amplification of 5 specific virulence factors found in CA-MRSA and HA-MRSA isolates. The specificity and precision of each method were evaluated using 237 clinical MRSA isolates. Results. The first method identified 100% and 93.2% of the CA-MRSA and HA-MRSA isolates, respectively. The second method also correctly identified the two isolates types (CA-MRSA and HA-MRSA). Conclusions. These two methods are a convenient alternative for the rapid identification of the CA-MRSA isolates, compared with other techniques such as pulsed field gel electrophoresis and multilocus sequence typing, which are time-consuming and more expensive.
Introducción. Los aislamientos de Staphylococcus aureus resistente a la meticilina asociado a la comunidad (SARM-AC), están aumentando la frecuencia de infecciones en personas sanas de la comunidad y en pacientes hospitalizados. En Colombia y en la región andina estos aislamientos tienen un componente genético relacionado con el clon pandémico USA300. Objetivo. Diseñar y estandarizar dos metodologías para la diferenciación rápida de aislamientos colombianos de S. aureus resistente a la meticilina asociado a la comunidad de los asociados al hospital (SARM-AH). Materiales y métodos. Se estandarizaron dos metodologías moleculares para la identificación de aislamientos de S. aureus resistente a la meticilina asociado a la comunidad. La primera se basa en la digestión diferencial con tres enzimas de restricción de los genes cinasa de carbamato (arcC)y cinasa de guanilato (gmk)para los tipos de secuencia 5 (ST5) y 8 (ST8), correspondientes a aislamientos de S. aureus resistente a la meticilina asociado al hospital y asociado a la comunidad, respectivamente. La segunda se basa en la amplificación por reacción en cadena de la polimerasa de cinco factores de virulencia que se encuentran de manera diferencial en estos aislamientos. Las dos metodologías fueron validadas en 237 aislamientos clínicos de S. aureus resistente a la meticilina. Resultados. Con la primera metodología se identificaron el 100 % y 93,2 % de los aislamientos de S. aureus resistente a la meticilina asociado a la comunidad y asociado al hospital, respectivamente. Con la segunda metodología se identificaron correctamente los dos tipos de aislamientos. Conclusiones. Estas dos metodologías son una buena alternativa en términos de ahorro en tiempo y dinero comparadas con otras técnicas, como la electroforesis en campo pulsado y la tipificación de secuencias multilocus para la rápida identificación de aislamientos de S. aureus resistente a la meticilina asociado a la comunidad en Colombia.
Assuntos
Humanos , Técnicas de Tipagem Bacteriana/métodos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Polimorfismo de Fragmento de Restrição , Reação em Cadeia da Polimerase/métodos , Infecções Estafilocócicas/microbiologia , Alelos , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana/normas , Colômbia , Infecções Comunitárias Adquiridas/epidemiologia , Infecções Comunitárias Adquiridas/microbiologia , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Eletroforese em Gel de Campo Pulsado , Genes Bacterianos , Guanilato Quinases/genética , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/genética , Fosfotransferases (Aceptor do Grupo Carboxila)/genética , Reprodutibilidade dos Testes , Alinhamento de Sequência , Análise de Sequência de DNA , Infecções Estafilocócicas/epidemiologia , Fatores de Tempo , Virulência/genéticaRESUMO
Introducción: el polimetilmetacrilato (PMMA) es el único sistema de entrega local de antibióticos aprobado por la FDA. Actualmente, este tipo de sistemas ha mejorado los resultados en el tratamiento de las infecciones asociadas con implantes ortopédicos con una seguridad biológica amplia, dada la disminución de toxicidad antibiótica sistémica. Se plantea la búsqueda de un sistema alternativo de entrega local de antibióticos debido a que en algunos casos el PMMA actúa como un cuerpo extraño, y en otros libera el antibiótico en rangos subterapéuticos actuando incluso como una superficie amigable para la formación de un biofilm por parte de las bacterias. Materiales y métodos: se diseñó un estudio in vitro, estandarizado, controlado, con el método de difusión agar, según los parámetros establecidos por el Clinical and Laboratory Standards Institute (CLSI) en el 2007 para determinar si el gel rico en plaquetas (PRP) suplementado con antibiótico sirve como sistema de transporte y liberación local de antibiótico inhibiendo el crecimiento de una cepa conocida de S. aureus. Resultados: los antibióticos evaluados fueron transportados y liberados por el gel rico en plaquetas. Se produjo inhibición del crecimiento bacteriano a mayores concentraciones de antibióticos. La oxacilina en la dilución 32 µg/ml tuvo un halo de inhibición mayor al sensidisco; la vancomicina generó los halos de inhibición de menor diámetro, y la ciprofloxacina presentó la menor capacidad de inhibición con respecto al control. Discusión: se demostró in vitro la capacidad que tiene el PRP como transportador y liberador de antibiótico. Dado el comportamiento heterogéneo recogido en los diferentes ensayos, se supone que cada antibiótico tiene una capacidad de difusión propia, la cual debe ser tenida en cuenta si se quiere emplear el gel de PRP como sistema de entrega local de antibióticos.
Assuntos
Antibacterianos , Plasma Rico em PlaquetasRESUMO
A comparative genetic analysis of 42 clinical Klebsiella pneumoniae isolates, resistant to two or more antibiotics belonging to the broad-spectrum ß-lactam group, sourced from Sydney, Australia, and three South American countries is presented. The study focuses on the genetic contexts of class 1 integrons, mobilizable genetic elements best known for their role in the rapid evolution of antibiotic resistance among Gram-negative pathogens. It was found that the class 1 integrons in this cohort were located in a number of different genetic contexts with clear regional differences. In Sydney, IS26-associated Tn21-like transposons on IncL/M plasmids contribute greatly to the dispersal of integron-associated multiple-drug-resistant (MDR) loci. In contrast, in the South American countries, Tn1696-like transposons on an IncA/C plasmid(s) appeared to be disseminating a characteristic MDR region. A range of mobile genetic elements is clearly being recruited by clinically important mobile class 1 integrons, and these elements appear to be becoming more common with time. This in turn is driving the evolution of complex and laterally mobile MDR units and may further complicate antibiotic therapy.
Assuntos
Farmacorresistência Bacteriana Múltipla/genética , Integrons/genética , Klebsiella pneumoniae/genética , Antibacterianos/farmacologia , Eletroforese em Gel de Campo Pulsado , Klebsiella pneumoniae/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Reação em Cadeia da PolimeraseRESUMO
Introducción. Staphylococcus aureus resistente a la meticilina (SARM) causa infecciones adquiridas en la comunidad y en el ámbito hospitalario. El ser portador de SARM se ha descrito como factor de riesgo para desarrollar infección clínica. Objetivo. Caracterizar la colonización por SARM en pacientes adultos de una unidad de cuidados intensivos colombiana, utilizando herramientas de biología molecular.Materiales y métodos. Entre febrero de 2007 y febrero de 2008 se tamizaron mediante hisopado nasofaríngeo, 705 pacientes al ingresar a la unidad de cuidados intensivos, de los cuales, 683 (96,9%) fueron seguidos semanalmente y al egreso de la unidad. Se determinó el perfil de sensibilidad de los aislamientos a 11 antibióticos por el método de dilución en agar; el 62,0% de los aislamientos de SARM fueron caracterizados genética y molecularmente. Resultados. Se tamizaron 705 pacientes al ingreso; 182 (25,8%) estaban colonizados por S. aureus, de los cuales, 51 (7,2%) eran resistentes a la meticilina. Se hizo el seguimiento durante la estancia en la unidad de cuidados intensivos a 683 pacientes, de los cuales, 62 (9,1%) fueron colonizados por SARM en dicha unidad. La prevalencia del clon chileno fue de 76,5% al ingreso y de 88,9% durante la estancia. El 16,0% de los pacientes colonizados desarrollaron algún tipo de infección por SARM. Se encontraron tres pacientes colonizados con SARM adquirido en la comunidad, los cuales fueron positivos para la leucocidina Panton-Valentine (Panton-Valentine leukocidin, PVL). Conclusiones. El 7,2% de los pacientes que ingresaron a la unidad de cuidados intensivos estaban colonizados con SARM. Éste es el primer reporte de colonización por aislamientos de SARM-ST8-SCCmec IVc adquirido en la comunidad y relacionado genéticamente con el clon pandémico USA300-0114 en Colombia.
Introduction. Methicillin-resistant Staphylococcus aureus (MRSA) cause nosocomial and community infections. MRSA colonization in hospitals has been described as an important risk factor during hospitalization. Objective. The colonization characteristics of MRSA was described using the tools of molecular biology. Materials and methods. Between February 2007 and February 2008, 705 patients entering a Colombian intensive care unit (ICU) were screened for MRSA by taking nasopharyngeal samples. For 683 of these patients, a weekly follow-up was provided after they left the ICU. The susceptibility of each S. aureus isolate was tested against 11 antibiotics using agar dilution methods. Sixty two percent (62.0%) of the MRSA isolates were characterized at genetic and molecular level with the detection of resistant genes, SCCmec typing using PCR and the genetic profile with pulsed field gel electrophoresis (PFGE). Results. Of the 705 patients screened at entry to the ICU, 182 (25.8%) were colonized by S. aureus, and of these, 51 (7.2%) were MRSA. Of the 683 patients with follow-up, 62 (9.1%) were infected by MRSA contracted in the hospital ICU. The prevalence of the Chilean clone was 76.5% at entry and 88.9% for follow-up patients. Of the 113 patients colonized with MRSA, nosocomial infection was present in 18 patients (16.0%). Three community-acquired MRSA isolates related to the USA300-0114 pandemic clone were identified. These were also positive for Panton-Valentine leucidin cytotoxin genes of S.aureus. Conclusions. This is the first report in Colombia of patients colonized with CA-MRSA-ST8-SCCmec IVc isolates, and it is a probable source of dissemination of this bacteria in Colombian hospitals.
Assuntos
Humanos , Cuidados Críticos , Staphylococcus aureus , Portador SadioRESUMO
BACKGROUND: Community-associated methicillin-resistant Staphylococcus aureus strains (CA-MRSA) have emerged as the causative agent of health care-associated infections. METHODS: An observational and prospective study was carried out in 5 hospitals in Bogotá, Colombia; severe MRSA infections were identified, and their origin led to classification as health care-associated (HA-MRSA), community-associated, or nosocomial infections. MRSA isolates were analyzed by SCCmec, pulsed-field gel electrophoresis, multilocus sequence typing, and virulence factors. RESULTS: Twenty-six (10.4%) CA-MRSA nosocomial infection-causing strains (eg, USA300) were detected in 250 MRSA infection isolates in mainly primary bacteremia and surgical site infections. The mortality related to nosocomial infection by CA-MRSA was 27%. CONCLUSION: The presence of nosocomial infection by CA-MRSA in Colombia was confirmed.