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2.
BMC Genomics ; 23(1): 20, 2022 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-34996351

RESUMO

BACKGROUND: Carbapenem-resistant hypervirulent K. pneumoniae (CR-hvKP) causes serious infections with significant morbidity and mortality. However, the epidemiology and transmission mechanisms of CR-hvKP and the corresponding carbapenem-resistant plasmids require further investigation. Herein, we have characterized an ST11 K. pneumoniae strain EBSI041 from the blood sample encoding both hypervirulence and carbapenem resistance phenotypes from a patient in Egypt. RESULTS: K. pneumoniae strain EBSI041 showed multidrug-resistance phenotypes, where it was highly resistant to almost all tested antibiotics including carbapenems. And hypervirulence phenotypes of EBSI041 was confirmed by the model of Galleria mellonella infection. Whole-genome sequencing analysis showed that the hybrid plasmid pEBSI041-1 carried a set of virulence factors rmpA, rmpA2, iucABCD and iutA, and six resistance genes aph(3')-VI, armA, msr(E), mph(E), qnrS, and sul2. Besides, blaOXA-48 and blaSHV-12 were harboured in a novel conjugative IncL-type plasmid pEBSI041-2. The blaKPC-2-carrying plasmid pEBSI041-3, a non-conjugative plasmid lacking the conjugative transfer genes, could be transferred with the help of pEBSI041-2, and the two plasmids could fuse into a new plasmid during co-transfer. Moreover, the emergence of the p16HN-263_KPC-like plasmids is likely due to the integration of pEBSI041-3 and pEBSI041-4 via IS26-mediated rearrangement. CONCLUSION: To the best of our knowledge, this is the first report on the complete genome sequence of KPC-2- and OXA-48-coproducing hypervirulent K. pneumoniae from Egypt. These results give new insights into the adaptation and evolution of K. pneumoniae during nosocomial infections.


Assuntos
Infecções por Klebsiella , Klebsiella pneumoniae , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Carbapenêmicos/farmacologia , Egito , Humanos , Infecções por Klebsiella/epidemiologia , Klebsiella pneumoniae/genética , Plasmídeos/genética , beta-Lactamases/genética
3.
mSphere ; 6(3)2021 05 19.
Artigo em Inglês | MEDLINE | ID: mdl-34011682

RESUMO

The emergence of carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates in Egyptian hospitals has been reported. However, the genetic basis and analysis of the plasmids associated with carbapenem-resistant hypervirulent K. pneumoniae (CR-HvKP) in Egypt have not been presented. Therefore, we attempted to decipher the plasmid sequences that are responsible for transferring the determinants of carbapenem resistance, particularly bla NDM-1 and bla KPC-2 Out of 34 K. pneumoniae isolates collected from two tertiary hospitals in Egypt, 31 were CRKP. Whole-genome sequencing revealed that our isolates were related to 13 different sequence types (STs). The most prevalent ST was ST101, followed by ST383 and ST11. Among the CRKP isolates, one isolate named EBSI036 has been reassessed by Nanopore sequencing. Genetic environment analysis showed that EBSI036 carried 20 antibiotic resistance genes and was identified as a CR-HvKP strain: it harbored four plasmids, namely, pEBSI036-1-NDM-VIR, pEBSI036-2-KPC, pEBSI036-3, and pEBSI036-4. The two carbapenemase genes bla NDM-1 and bla KPC-2 were located on plasmids pEBSI036-1-NDM-VIR and pEBSI036-2-KPC, respectively. The IncFIB:IncHI1B hybrid plasmid pEBSI036-1-NDM-VIR also carried some virulence factors, including the regulator of the mucoid phenotype (rmpA), the regulator of mucoid phenotype 2 (rmpA2), and aerobactin (iucABCD and iutA). Thus, we set out in this study to analyze in depth the genetic basis of the pEBSI036-1-NDM-VIR and pEBSI036-2-KPC plasmids. We report a high-risk clone ST11 KL47 serotype of a CR-HvKP strain isolated from the blood of a 60-year-old hospitalized female patient from the intensive care unit (ICU) in a tertiary care hospital in Egypt, which showed the cohabitation of a novel hybrid plasmid coharboring the bla NDM-1 and virulence genes and a bla KPC-2-carrying plasmid.IMPORTANCE CRKP has been registered in the critical priority tier by the World Health Organization and has become a significant menace to public health. The emergence of CR-HvKP is of great concern in terms of both disease and treatment. In-depth analysis of the carbapenemase-encoding and virulence plasmids may provide insight into ongoing recombination and evolution of virulence and multidrug resistance in K. pneumoniae Thus, this study serves to alert contagious disease clinicians to the presence of hypervirulence in CRKP isolates in Egyptian hospitals.


Assuntos
Antibacterianos/farmacologia , Carbapenêmicos/farmacologia , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/patogenicidade , Plasmídeos/genética , Fatores de Virulência/genética , beta-Lactamases/genética , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Egito , Feminino , Humanos , Lactente , Recém-Nascido , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/genética , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Centros de Atenção Terciária/estatística & dados numéricos , Adulto Jovem
4.
Photodiagnosis Photodyn Ther ; 34: 102301, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33894372

RESUMO

BACKGROUND: The growing emergence of microbial resistance to antibiotics represents a worldwide challenge. Antimicrobial photodynamic inactivation (aPDI) has been introduced as an alternative technique, especially when combined with nanotechnology. Therefore, this study was designed to investigate the therapeutic merits of combined aPDI and nanoemulsion in infections caused by resistant bacterial strains. METHODS: Cationic zinc (II) phthalocyanine nanoemulsions (ZnPc-NE) were prepared using isopropyl myristate (IPM) as oil phase, egg phosphatidylcholine (egg PC) as emulsifier, and N-cetyl-N,N,N-trimethyl ammonium bromide (CTAB). Nanoemulsions were characterized for particle size, polydispersity, zeta potential, viscosity, and skin deposition. The in-vitro aPDI was investigated on human resistant pathogens; gram-positive methicillin-resistant Staphylococcus aureus (MRSA) and gram-negative Multidrug-resistant strain of Escherichia coli (MDR E. coli), under different experimental conditions. In addition, in-vivo model of abrasion wound infected by MDR E. coli was induced in rats to investigate the therapeutic potential of the selected formulation. RESULTS: It was evident that the selected ZnPc formulation (20 % IPM, 2 % egg PC and 0.5 % CTAB) displayed a particle size of 209.9 nm, zeta potential +73.1 mV, and 23.66 % deposition of ZnPc in skin layers. Furthermore, the selected formulation combined with light achieved almost 100 % eradication of the two bacterial strains, with superior bacterial load reduction and wound healing propertiesin-vivo, compared to either the nanoemulsion formulation or laser alone. CONCLUSION: ZnPc nanoemulsion improved antimicrobial photodynamic therapy in inactivating resistant bacterial infections and provided a promising therapeutic means of treating serious infections, and hence could be applied in diseases caused by other bacterial strains.


Assuntos
Staphylococcus aureus Resistente à Meticilina , Fotoquimioterapia , Animais , Antibacterianos/farmacologia , Escherichia coli , Indóis , Isoindóis , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Ratos , Zinco
5.
Sci Rep ; 10(1): 5156, 2020 03 20.
Artigo em Inglês | MEDLINE | ID: mdl-32198440

RESUMO

Hepatocellular carcinoma (HCC) is the second most common cause of cancer-related death worldwide. The use of alpha fetoprotein (AFP) alone was not an accurate biomarker for HCC despite its high specificity. Therefore, we assessed the possible role of serum biomarkers that have been mentioned briefly in previous studies on Egyptian patients ion top of HCV. However these studies included small number of patients and did not assess the different stages of hepatocarcinogenesis. In the current study we assessed 1) the expression levels of Golgi protein 37(GP73),Midkine (MDK) and Dickkopf-1(DKK-1) proteins separately and in combination at different stages of hepatocarcinogenesis. GP73, MDK and DKK-1 proteins were assessed in 238 individuals divided into 4 groups (HCC, chronic HCV, and chronic HCV with cirrhosis and healthy subjects as a control) Serum levels of GP73, MDK, and DKK-1 were assessed in all subjects by ELISA. Serum levels of the studied markers were significantly higher in HCC compared to other groups (p < 0.001). The ROC curve analysis for the studied markers showed 1) 88.5% sensitivity, 80.6% specificity, 69% PPV, 93.5% NPV and (AUC 0.91)for MDK; 2) 93.6%, 86.9%, 77.7%, 96.5% for DKK-1. 3) 91%, 85%, 74.7%, 95% (AUC 0.96) for GP73 and 4) 74.4%, 84.4%, 69.9%, 87.1% (AUC 0.81) for AFP. Serum levels of GP73, MDK, and DKK-1 are comparable to AFP as promising predictor biomarkers for HCC patients from Egypt. A two markers panel including Gp73 and DKK-1 showed the highest specificity and sensitivity among different markers combinations.


Assuntos
Carcinoma Hepatocelular/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Neoplasias Hepáticas/metabolismo , Proteínas de Membrana/análise , Midkina/metabolismo , Adulto , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/sangue , Carcinoma Hepatocelular/patologia , Feminino , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/sangue , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas/patologia , Masculino , Proteínas de Membrana/sangue , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Midkina/sangue , Prognóstico , Precursores de Proteínas/sangue , Curva ROC , Sensibilidade e Especificidade , alfa-Fetoproteínas/metabolismo
6.
J Glob Antimicrob Resist ; 12: 68-72, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29274469

RESUMO

OBJECTIVES: The incidence of reduced susceptibility to tigecycline (TIG) is increasing. This study aimed to analyse the in vitro activity of TIG against Enterococcus spp. isolates recovered from hospitalised patients and to evaluate the effect of omeprazole on the in vitro antimicrobial activity of TIG against several enterococcal species. METHODS: A total of 67 Enterococcus clinical isolates were identified by MALDI-TOF/MS and multiplex PCR. Minimum inhibitory concentrations (MICs) of TIG alone and in combination with omeprazole (10, 30 and 60mg/L) were determined by broth microdilution. Antibiotic susceptibility to other antibiotics was determined by disk diffusion. The presence of van, tet(X) and tet(X1) genes was tested by multiplex PCR. RESULTS: Of the 67 Enterococcus isolates, 2 (3.0%) were resistant to TIG and 13 (19.4%) were intermediate-resistant according to EUCAST. The frequencies of resistance to norfloxacin (80.6%), doxycycline (80.6%), levofloxacin (74.6%) and ciprofloxacin (71.6%) were highest, whilst that of vancomycin (25.4%) was lowest. The vanA gene was detected in 11 Enterococcus isolates (8 Enterococcus faecalis, 3 Enterococcus faecium), vanB in 3 Enterococcus isolates (2 E. faecium, 1 E. faecalis) and vanC-2/3 in 3 Enterococcus casseliflavus. Nine isolates (13.4%) were positive for tet(X1). TIG resistance occurred both in patients receiving or not TIG and/or omeprazole. Omeprazole increased TIG MICs by 4-128-fold. CONCLUSIONS: The possibility of selection of TIG-non-susceptible Enterococcus in the gut may occur with long-term use of omeprazole. Omeprazole influenced TIG activity in a concentration-dependent manner. To our knowledge; this is the first report of TIG-non-susceptible Enterococcus spp. in Egypt.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecium/efeitos dos fármacos , Infecções por Bactérias Gram-Positivas/microbiologia , Inibidores da Bomba de Prótons/farmacologia , Tigeciclina/farmacologia , Adolescente , Adulto , Aspartato Aminotransferases/sangue , Egito/epidemiologia , Enterococcus faecalis/isolamento & purificação , Enterococcus faecium/isolamento & purificação , Feminino , Genes Bacterianos , Infecções por Bactérias Gram-Positivas/epidemiologia , Hospitais Universitários , Humanos , Incidência , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Omeprazol/farmacologia , Reação em Cadeia da Polimerase , Adulto Jovem
7.
Microb Drug Resist ; 24(3): 269-277, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28783427

RESUMO

Of great concern is the increased frequency of carbapenem-resistant Acinetobacter baumannii (CRAB) causing healthcare-associated infections. Different classes of ß-lactamases are involved in this resistance through hydrolyzing carbapenems. Multilocus sequence typing (MLST) has been applied successfully for characterizing different varieties of bacterial pathogens epidemiologically. In the present study, we aimed to type and characterize the resistance profile of clinical isolates of CRAB causing healthcare-associated infections in patients admitted to Kasr Al-Aini hospital, using MLST, and compare with sequence types (STs) from other countries. A total of 50 isolates were collected from clinical samples (predominantly wound and blood), then identified by blaOXA-51-like gene PCR, and subjected to Oxford MLST scheme. The ST was designated according to PubMLST database, and e-BURST algorithm was used to assign clonal complexes. Four sets of multiplex PCR were performed to detect common carbapenem resistance genes. ST391 was the predominant ST detected in 17 cases, 70.5% of which harbored blaOXA-23 alone, both blaOXA-23 and blaKPC in 11.8%. Newly recognized 13 STs were submitted to the PubMLST database. Carbapenem resistance due to blaOXA-23 carbapenemase was detected in 36/50 (72%), followed by blaOXA-23 concomitant with blaKPC in 7/50 (14%), while blaNDM with blaOXA-58 in 3/50 (6%) and blaNDM alone in 1 case (2%). To conclude, this study demonstrates the propagation of highly resistant clone of STs 391 and 1151, carrying blaOXA-23 genes, with the first report of blaKPC in blaOXA carrying CRAB and the presence of new STs by performing the MLST technique in an Egyptian laboratory facility.


Assuntos
Infecções por Acinetobacter/epidemiologia , Acinetobacter baumannii/genética , Proteínas de Bactérias/genética , Infecção Hospitalar/epidemiologia , Farmacorresistência Bacteriana Múltipla/genética , Resistência beta-Lactâmica/genética , beta-Lactamases/genética , Infecções por Acinetobacter/tratamento farmacológico , Infecções por Acinetobacter/microbiologia , Acinetobacter baumannii/classificação , Acinetobacter baumannii/efeitos dos fármacos , Acinetobacter baumannii/isolamento & purificação , Adulto , Idoso , Antibacterianos/farmacologia , Carbapenêmicos/farmacologia , Células Clonais , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/microbiologia , Egito/epidemiologia , Feminino , Expressão Gênica , Genótipo , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Epidemiologia Molecular , Tipagem de Sequências Multilocus , Centros de Atenção Terciária
8.
J Microbiol ; 54(1): 39-43, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26727900

RESUMO

Endoscopes are a common source of outbreaks of healthcare-associated infections. It is therefore important to identify high-level disinfectants capable of eliminating or killing all vegetative bacteria, mycobacteria, and viruses. Aldehydebased disinfectants are most commonly used in clinical practice but resistance has recently been detected and side effects associated with these disinfectants are well documented. In this study, we evaluated Virusolve+® EDS, a novel quaternary ammonium compound formulation supplied by Amity international, against Mycobacterium bovis (ATCC-27289), hepatitis C virus (HCV)-positive serum and hepatitis B surface antigen-positive serum. We also compared its efficacy against Cidex® (glutaraldehyde 2%), an aldehyde-based disinfectant. M. bovis showed no growth after 10 weeks with either Virusolve+® or Cidex®. Virusolve+® achieved a 10(4)- fold reduction in the initial 10(6) HCV load under clean conditions (without red blood cells) for 20 min, whereas Cidex® achieved this reduction under clean and dirty conditions (without and with red blood cells, respectively) after both 10 and 20 min. Both Virusolve+® and Cidex® were able to eradicate hepatitis B virus (HBV) infectivity under clean conditions after 10 and 20 min, whereas under dirty conditions they were only able to eradicate virus infectivity after 20 min. Virusolve+® EDS when compared with Cidex® showed equal mycobactericidal activity completely eradicating M. bovis. However, both showed comparable virucidal activity against HBV, which was more effective under clean conditions, emphasizing the importance of the cleaning step in endoscope reprocessing. Cidex® was more effective at eradicating HCV under dirty conditions after a short contact time.


Assuntos
Desinfetantes/farmacologia , Hepacivirus/crescimento & desenvolvimento , Vírus da Hepatite B/crescimento & desenvolvimento , Mycobacterium bovis/crescimento & desenvolvimento , Compostos de Amônio Quaternário/farmacologia , Glutaral , Hepacivirus/efeitos dos fármacos , Vírus da Hepatite B/efeitos dos fármacos , Humanos , Mycobacterium bovis/efeitos dos fármacos , Fatores de Tempo , Carga Viral , Viremia
9.
Ann Saudi Med ; 35(6): 440-4, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26657227

RESUMO

BACKGROUND: The rapid and accurate identification of nosocomial clinical isolates is the first essential step in investigating nosocomial outbreaks. We aimed to evaluate the performance of MDR-CHROMagar Acinetobacter versus matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) in rapid detection of nosocomial Acinetobacter baumannii isolated from patients admitted to the surgical intensive care unit (SICU) of Kasr Alainy- Cairo University. METHODS: Over a period of 9 months from January 2014 until September 2014, 234 samples were collected. All samples were directly cultured on MDR-CHROMagar Acinetobacter media. MALDI-TOF MS was used to identify all non-lactose fermenting colonies on conventional media. Confirmation of species identification was done by detecting the blaOXA-51 like gene by PCR. RESULTS: Statistical evaluation of MDR-CHROMagar Acinetobacter against blaOXA-51 like PCR as the reference method for identification of A baumannii showed a sensitivity of 100% (95% confidence interval [CI]: 93.36% to 100%), specificity 98.8% (95% confidence interval [CI]: 96.04% to 99.68%), positive predictive value 96.4% (95%CI: 86.61% to 99.37%), negative predictive value 100% (95% CI: 97.36% to 100%). The statistical evaluation of MALDI-TOF against blaOXA-51 PCR was 100% concordance. CONCLUSION: MALDI-TOF MS was more specific than CHROMagar in identifying Acinetobacter spp and allowed further identification of non-A Baumannii species such as A hemolyticus and A nosocomialis, which are less common Acinetobacter spp involved in hospital-acquired infections.


Assuntos
Infecções por Acinetobacter/diagnóstico , Acinetobacter baumannii/isolamento & purificação , Técnicas Bacteriológicas/métodos , Infecção Hospitalar/diagnóstico , Infecções por Acinetobacter/microbiologia , Infecção Hospitalar/microbiologia , Egito , Humanos , Unidades de Terapia Intensiva , Reação em Cadeia da Polimerase , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
10.
J Infect Dev Ctries ; 8(10): 1252-8, 2014 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-25313600

RESUMO

INTRODUCTION: Diagnosis of bloodstream infections using bacteriological cultures suffers from low sensitivity and reporting delay. Advanced molecular techniques introduced in many laboratories provide rapid results and may show improvements in patient outcomes. This study aimed to evaluate the usefulness of a molecular technique, broad-range 16S rRNA PCR followed by sequencing for the diagnosis of bloodstream infections, compared to blood culture in different patient groups. METHODOLOGY: Conventional PCR was performed, using broad-range 16S rRNA primers, on blood cultures collected from different patients with suspected bloodstream infections; results were compared with those of blood culture. RESULTS: Though blood culture is regarded as the gold standard, PCR evaluation showed sensitivity of 86.25%, specificity of 91.25%, positive predictive value of 76.67%, negative predictive value of 95.22%, and accuracy of 88.8%. CONCLUSIONS: Molecular assays seem not to be sufficient to replace microbial cultures in the diagnosis of bloodstream infections, but they can offer a rapid, good negative test to rule out infection due to their high negative predictive value.


Assuntos
Bacteriemia/diagnóstico , Bacteriemia/microbiologia , Bactérias/isolamento & purificação , Técnicas Bacteriológicas/métodos , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 16S/genética , Bactérias/classificação , Bactérias/genética , Humanos , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Análise de Sequência de DNA/métodos , Fatores de Tempo
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