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2.
Folia Microbiol (Praha) ; 56(2): 122-30, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21431912

RESUMO

Polymerase chain reaction (PCR) amplification of antibiotic resistance genes as well as staphylococcal cassette chromosome mec (SCCmec) typing and pulsed-field gel electrophoresis (PFGE) of SmaI macrorestriction fragments of genomic DNA were used to characterize 45 methicillin-resistant coagulase-negative staphylococci (MRCoNS) isolates responsible of bacteremia recovered in patients at the Bone Marrow Transplant Centre of Tunisia in 1998-2007. Among the 45 MRCoNS isolates, Staphylococcus epidermidis was the most prevalent species (75.6%) followed by Staphylococcus haemolyticus (22.2%) and Staphylococcus hominis (2.2%). Extended susceptibility profiles were generated for MRCoNS against 16 antimicrobial agents. Out of 45 mecA-positive strains, 43 (95.6%) were phenotypically methicillin-resistant and two (4.4%) were methicillin-susceptible. The msr(A) was the most prevalent gene (13 isolates; 48.1%) among erythromycin-resistant isolates. The erm(C) was found alone in seven (25.9%) or in combination with both erm(A) and erm(B) in two (7.4%) isolates. The aac(6')-Ie-aph(2″)-Ia was the most prevalent gene among aminoglycoside-resistant isolates, detected alone in 14 isolates (33.3%) isolates, in combination with ant(4')-Ia in 18 (42.8%) isolates, in combination with aph(3')-IIIa in four (9.5%) or with both ant(4')-Ia and aph(3')-IIIa in two (4.7%) isolates. The ant(4')-Ia was detected in three (7.1%) isolates and the aph(3')-IIIa in one (2.4%) isolate. Among tetracycline-resistant isolates, six (85.7%) strains harbored the tet(K) gene and one (14.3%) strain carried tet(K) and tet(M) genes. SCCmec types IV (31%) and III (24.5%), the most prevalent types detected, were found to be more resistant to non-ß-lactam antibiotics. A wide diversity of isolates was observed by PFGE among MRCoNS.


Assuntos
Bacteriemia/microbiologia , Resistência a Meticilina , Infecções Estafilocócicas/microbiologia , Staphylococcus epidermidis/isolamento & purificação , Staphylococcus haemolyticus/isolamento & purificação , Staphylococcus hominis/isolamento & purificação , Antibacterianos/farmacologia , Técnicas de Tipagem Bacteriana , Coagulase/metabolismo , DNA Bacteriano/genética , DNA Bacteriano/metabolismo , Desoxirribonucleases de Sítio Específico do Tipo II/metabolismo , Farmacorresistência Bacteriana , Eletroforese em Gel de Campo Pulsado , Genes Bacterianos , Neoplasias Hematológicas/complicações , Humanos , Testes de Sensibilidade Microbiana , Tipagem Molecular , Reação em Cadeia da Polimerase/métodos , Staphylococcus epidermidis/classificação , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/genética , Staphylococcus haemolyticus/classificação , Staphylococcus haemolyticus/efeitos dos fármacos , Staphylococcus haemolyticus/genética , Staphylococcus hominis/classificação , Staphylococcus hominis/efeitos dos fármacos , Staphylococcus hominis/genética , Tunísia
3.
Pathol Biol (Paris) ; 59(4): 199-206, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19481372

RESUMO

To investigate the prevalence of resistance to macrolide, lincosamide and streptogramin (MLS) antibiotics in Gram-positive cocci isolated in a Bone Marrow Transplant Center of Tunisia, we tested the antibiotic susceptibility of 172 clinical isolates of Staphylococcus epidermidis, Streptococcus mitis and Enterococcus faecium to macrolide erythromycin and spiramycin, the lincosamide clindamycin and the streptogramin pristinamycin. These three groups of organisms were mostly resistant to macrolides and lincosamide, but were commonly susceptible to pristinamycin. The resistance phenotypes of erythromycin-resistant isolates were determined by the five-disc test with erythromycin, spiramycin, lincomycin, clindamycin and pristinamycin, which showed that most exhibited constitutive MLS resistance. In order to determine the prevalence of the resistance genotypes and the resistance mechanisms, the prevalence of the erythromycin resistance methylase (erm) (A), erm(B), erm(C), msr(A) and macrolide efflux (mef) (A) genes in the erythromycin-resistant isolates was identified by polymerase chain reaction (PCR) analysis. The resistance was due mainly to the presence of ermB in E. faecium (80%), ermC in S. epidermidis (53%) and mefA in S. mitis (65%).


Assuntos
Transplante de Medula Óssea , Farmacorresistência Bacteriana , Cocos Gram-Positivos/efeitos dos fármacos , Lincosamidas/farmacologia , Macrolídeos/farmacologia , Estreptograminas/farmacologia , DNA Bacteriano/análise , Farmacorresistência Bacteriana/genética , Enterococcus faecium/efeitos dos fármacos , Genótipo , Cocos Gram-Positivos/genética , Humanos , Testes de Sensibilidade Microbiana , Fenótipo , Staphylococcus epidermidis/efeitos dos fármacos , Streptococcus mitis/efeitos dos fármacos , Tunísia
4.
Curr Microbiol ; 59(4): 380-5, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19588195

RESUMO

Staphylococcal Cassette Chromosome mec (SCCmec) is a mobile genetic element that carries the gene mecA mediating the methicillin resistance in staphylococci. It is composed of mec and ccr gene complexes. Six SCCmec types have been defined so far. SCCmec typing of 13 methicillin-resistant Staphylococcus aureus (MRSA) out of 72 (18%) non redundant S. aureus strains recovered in 1998-2007 at the Bone Marrow Transplant Centre of Tunis was carried out. The isolates were identified by conventional methods. Antibiotic susceptibility was determined by oxacillin and cefoxitin disks and oxacillin MIC by E-test. Methicillin resistance was detected by mecA PCR. The SCCmec complex types were determined by PCR. The epidemiology of MRSA has been investigated by PFGE. Among 13 mecA positive strains, 12 were resistant to oxacillin (MIC = 3 to >256 microg/microl) and to cefoxitin and one strain was pre-resistant: susceptible to oxacillin (MIC = 0.19 microg/microl) and to cefoxitin. Hospital-acquired MRSA (HA-MRSA) strains had essentially SCCmec type IV (nine strains) or III (two strains) or I (one strain). One strain shown to carry ccrAB1 and ccrAB2 genes in combination with class B mec. Seven of 13 MRSA strains isolated from 2000 to 2006 were classified with major similarity group A harbored SCCmec type IV.


Assuntos
Técnicas de Tipagem Bacteriana , Cromossomos Bacterianos , Infecção Hospitalar/microbiologia , DNA Bacteriano/genética , Staphylococcus aureus Resistente à Meticilina/classificação , Reação em Cadeia da Polimerase/métodos , Infecções Estafilocócicas/microbiologia , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Análise por Conglomerados , Eletroforese em Gel de Campo Pulsado , Genótipo , Hospitais , Humanos , Sequências Repetitivas Dispersas , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Testes de Sensibilidade Microbiana , Epidemiologia Molecular , Proteínas de Ligação às Penicilinas , Tunísia , beta-Lactamas/farmacologia
5.
Pathol Biol (Paris) ; 57(5): 420-4, 2009 Jul.
Artigo em Francês | MEDLINE | ID: mdl-18838231

RESUMO

We propose a simple and rapid method to discriminate SHV-type extended spectrum beta-lactamase (ESBL) genes in P. aeruginosa based on PCR techniques (PCR-RFLP and RSI-PCR). We studied 22 producing ESBL P. aeruginosa strains isolated from seven immunocompromised patients (19 isolates) and from environmental swabs (three isolates) at the Bone Marrow Transplantation Center of Tunis. Screening PCR with primer pairs designed to detect gene encoding TEM, SHV, OXA group I, OXA group II, OXA-18 and PER-1 ESBL was positive for bla(OXA18) and bla(SHV) genes in all isolates. Pulsed field gel electrophoresis using SpeI endonuclease defined five genotypic groups. For at least one isolate corresponding to each genotype observed, restriction of PCR products by DdeI and BsrI revealed the same restriction pattern that the bla(SHV-1) negative control; in the same way, RSI-PCR products digestion by NruI, thus excluding 35, 238 and 240 mutations characterizing reported ESBL in P. aeruginosa (SHV-2a, SHV5 et SHV12), and suggesting that studied bla(SHV) genes were not ESBL ones. Genomic DNA hybridization by southern blot with probe consisting in bla(SHV-1) gene was positive in these isolates. Sequencing the full-length open reading frame revealed nucleotide sequence of the bla(SHV-1). PCR-RFLP and RSI-PCR results were then confirmed. This approach is effective for screening P. aeruginosa for ESBL genes carriage in epidemiological studies and for detecting new variants.


Assuntos
Proteínas de Bactérias/genética , Genes Bacterianos , Reação em Cadeia da Polimerase/métodos , Pseudomonas aeruginosa/genética , Resistência beta-Lactâmica/genética , beta-Lactamases/genética , Proteínas de Bactérias/classificação , Southern Blotting , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Contaminação de Equipamentos , Humanos , Hospedeiro Imunocomprometido , Programas de Rastreamento , Mutagênese Insercional , Polimorfismo de Fragmento de Restrição , Infecções por Pseudomonas/epidemiologia , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/enzimologia , Especificidade por Substrato , Tunísia/epidemiologia , beta-Lactamases/classificação
6.
Pathol Biol (Paris) ; 57(5): 363-7, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18178029

RESUMO

OBJECTIVE: The aim of this study was to investigate the molecular epidemiology of Stenotrophomonas maltophilia strains responsible for respiratory infection in a neonatal intensive care unit (NICU) in Tunis City, isolated during 22 months (December 2003-September 2005). MATERIALS AND METHODS: Twelve strains of S. maltophilia isolated from tracheal aspirates of distinct infants and two environmental strains were tested for antibiotic susceptibility and genotyped by pulsed-field gel electrophoresis (PFGE) method. RESULTS: Unlike a large heterogeneity demonstrated by the antibiotyping method, PFGE identified two concomitant outbreaks consisting of nine, including an environmental strain (clone A), and four strains (clone B), respectively; a distinguishable strain was classified in a unique pattern (PFGE type C). The long-term dissemination of these strains is a characteristic feature of these outbreaks. Improvement of hygienic conditions attributed to a markedly decrease in their isolation frequencies. Concomitant outbreaks and long period persistence of S. maltophilia in NICU is an important finding of this study. CONCLUSION: Identification of two clonal strains of S. maltophilia responsible of respiratory infection. Epidemic strains are hardly eradicated when colonization is established.


Assuntos
Infecção Hospitalar/microbiologia , Surtos de Doenças , Infecções por Bactérias Gram-Negativas/microbiologia , Unidades de Terapia Intensiva Neonatal , Stenotrophomonas maltophilia/isolamento & purificação , Traqueíte/microbiologia , Infecção Hospitalar/epidemiologia , DNA Bacteriano/análise , Farmacorresistência Bacteriana Múltipla , Eletroforese em Gel de Campo Pulsado , Genótipo , Infecções por Bactérias Gram-Negativas/epidemiologia , Humanos , Recém-Nascido , Stenotrophomonas maltophilia/classificação , Stenotrophomonas maltophilia/efeitos dos fármacos , Stenotrophomonas maltophilia/genética , Traqueia/microbiologia , Traqueíte/epidemiologia , Tunísia
7.
Pathol Biol (Paris) ; 57(5): 353-7, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18178031

RESUMO

OBJECTIVE: The aim of this study was to determine the overall percentage of beta-lactams susceptibility, beta-lactamase production, penicillin binding protein (PBP) modification and serotypes of colonizing Haemophilus influenzae strains. DESIGN: A total of 50 isolates of colonized H. influenzae, isolated from neutropenic patients. The prevalence of beta-lactams resistance and beta-lactamase production were recorded for each strains using E-test strips and chromogenic cephalosporin test, then were determined their resistance genes (bla(TEM) and bla(ROB)) by PCR as well as their capsular types by standard slide agglutination serotyping (SAST) and capsular genes amplification. RESULTS: Thirty-two percent of the 50 strains were amoxicillin resistant, among these, 20% were resistant by beta-lactamase production, and they produced all type TEM beta-lactamase. Four percent of the isolates had PBP modification and three strains (6%) associated the two resistance mechanisms. Slide agglutination serotyping showed that 95.8% of the strains were unencapsulated, and 4.1% were of serogroup b. The result was confirmed by PCR capsular typing. CONCLUSION: By the light of these results, our findings suggest that it becomes important to follow the evolution of the resistance background of our strains, and that the majority of colonizing H. influenzae strains isolated in our center are unencapsulated.


Assuntos
Cápsulas Bacterianas/química , Portador Sadio/microbiologia , Farmacorresistência Bacteriana Múltipla/genética , Infecções por Haemophilus/microbiologia , Haemophilus influenzae/classificação , Neutropenia/microbiologia , Resistência beta-Lactâmica/genética , Técnicas de Tipagem Bacteriana , Transplante de Medula Óssea , Portador Sadio/epidemiologia , DNA Bacteriano/genética , Infecções por Haemophilus/epidemiologia , Haemophilus influenzae/química , Haemophilus influenzae/genética , Haemophilus influenzae/isolamento & purificação , Humanos , Hospedeiro Imunocomprometido , Testes de Sensibilidade Microbiana , Fenótipo , Sorotipagem , Tunísia/epidemiologia , beta-Lactamases/genética
8.
Pathol Biol (Paris) ; 57(3): 268-71, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-18394823

RESUMO

OBJECTIVES: Investigation of the occurrence and antibiotic susceptibility of Enterococcus faecium isolates, collected during four years from neutropenic patients at the Tunisian bone marrow transplantation centre. MATERIALS AND METHODS: E. faecium strains were identified by conventional methods and by the Api20 Strep (Bio-Mérieux, France). Antibiotic susceptibility was determined by the disk diffusion method on Mueller-Hinton agar and interpreted as recommended by CA-SFM. MICs of ampicillin, vancomycin, and teicoplanin were determined by E-test method. RESULTS: Two hundred and thirty five E. faecium isolates were recovered from stool cultures or rectal swabs (229), throat (three), urine (two), and pus of wound (one). None was responsible for bacteraemia. Ampicillin resistance, without production of beta-lactamase, was observed in 43.8% of isolates. All the isolates were susceptible to glycopeptides. High rates of resistance were observed: high-level resistance (HLR) to gentamicin (33.6%), HLR to kanamycin (55.7%), HLR to streptomycin (47.6%), erythromycin (86.4%), ciprofloxacin (78.7%), rifampicin (85%), and tetracycline (43%). Strains with HLR to gentamicin were significantly more resistant to ampicillin and streptomycin. Multiple drug resistance was observed in most isolates. CONCLUSION: These findings demonstrated the low pathogenic power of E. faecium in our patients, and the high frequencies of resistance to ampicillin and aminoglycosides. In the absence of glycopeptide-resistance, vancomycin remains an alternative treatment against multidrug resistant strains.


Assuntos
Infecções Bacterianas/tratamento farmacológico , Transplante de Medula Óssea/efeitos adversos , Enterococcus faecium/isolamento & purificação , Infecções por Bactérias Gram-Positivas/epidemiologia , Infecções por Bactérias Gram-Positivas/transmissão , Resistência a Ampicilina , Antibacterianos/farmacologia , Infecções Bacterianas/epidemiologia , Resistência Microbiana a Medicamentos , Resistência a Múltiplos Medicamentos , Enterococcus faecium/efeitos dos fármacos , Fezes/microbiologia , Glicopeptídeos/farmacologia , Humanos , Complicações Pós-Operatórias/microbiologia , Tunísia/epidemiologia
9.
Pathol Biol (Paris) ; 57(3): e73-5, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-18456431

RESUMO

Twelve multidrug-resistant Pseudomonas aeruginosa (MDRPA) isolates were recovered over a period of two years in the National Bone Marrow Transplant Centre of Tunisia. MDRPA isolates were isolated from seven patients and from three environmental samples. Isoelectric focusing revealed pIs of 8.2, 5.5 and 7.6 in all MDRPA isolates. These strains produced the OXA-18 extended spectrum beta-lactamase and an SHV type beta-lactamase as shown by screening PCR analysis. DNA hybridization confirmed this inference, detecting bla(SHV) gene in these isolates. Pulsed-field gel electrophoresis (PFGE) defined one predominant genomic group; group A (seven isolates) and four different genotypes containing one to two isolates. Clonally related isolates were recovered from three patients and from two washbasins. Sequencing DNA of cluster representative strains identified the classical bla(SHV-1) gene. For these strains, the nucleotide sequence of the structural bla(SHV-1) gene was nearly identical to those previously described. Such enzyme has not been reported from P. aeruginosa. This is the first report of the SHV-1 penicillinase in epidemic P. aeruginosa strain.


Assuntos
Pseudomonas aeruginosa/genética , beta-Lactamases/genética , Primers do DNA , DNA Bacteriano/genética , Farmacorresistência Bacteriana , Resistência a Múltiplos Medicamentos , Eletroforese em Gel de Campo Pulsado , Enterobacteriaceae/enzimologia , Humanos , Focalização Isoelétrica , Klebsiella/enzimologia , Testes de Sensibilidade Microbiana , Penicilinase/genética , Penicilinase/isolamento & purificação , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/enzimologia , Pseudomonas aeruginosa/isolamento & purificação , beta-Lactamases/isolamento & purificação , beta-Lactamases/metabolismo
10.
Lett Appl Microbiol ; 46(3): 358-63, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18194160

RESUMO

AIMS: We report the analysis of genetic determinants conferring resistance to pristinamycin in Staphylococcus epidermidis strains and epidemiology typing of these strains by pulsed-field gel electrophoresis. METHODS AND RESULTS: Staphylococcus epidermidis (346 isolates) were searched for strains with pristinamycin resistance. Pristinamycin-resistant strains (seven isolates) were isolated in five patients with haematological cancer in the Bone Marrow Transplant Centre of Tunisia in 2002. Resistance to pristinamycin was observed in 2% of isolates. The seven pristinamycin-resistant strains shared resistance to oxacillin (MIC = 8-512 microg ml(-1)), gentamicin (MIC = 16-512 microg ml(-1)), erythromycin (MIC > 1024 microg ml(-1)), lincomycin (MIC > 1024 microg ml(-1)), pristinamycin (MIC = 4-16 microg ml(-1)) and rifampin (MIC = 128-256 microg ml(-1)). erm genes were amplified: ermA from six strains and ermC from one. vga gene encoding streptogramins A resistance (pristinamycin résistance) was amplified from all strains and typed as vgaA by analysis after electrophoresis of restriction profiles of vga amplicons (two fragments with Sau3A of 164 and 378 bp; one fragment with EcoRI). Pulsed-field gel electrophoresis (PFGE) of SmaI chromosomal DNA digests of the seven S. epidermidis isolates divided them into two distinct pattern types: pulsed-field type A (classified from A1 to A6 subtypes) and type B. The six strains harbouring ermA genes belonged to the PFGE type A while the strain harbouring ermC genes belonged to the PFGE type B. We characterized an epidemic strain carrying the vgaA and ermA genes responsible for the outbreak. CONCLUSIONS: Two clones of pristinamycin-resistant S. epidermidis were isolated in our patients. One of them, isolated in all patients, had expanded over six months suggesting acquisition by cross-contamination. SIGNIFICANCE AND IMPACT OF THE STUDY: Increasing isolation of pristinamycin resistant S. epidermidis strains is an alarming indicator of nosocomial dissemination. The vector will be determined to establish a system of epidemiological surveillance.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Neoplasias Hematológicas/complicações , Pristinamicina/farmacologia , Infecções Estafilocócicas/epidemiologia , Staphylococcus epidermidis/efeitos dos fármacos , Proteínas de Bactérias/genética , Transplante de Medula Óssea , Primers do DNA , Eletroforese em Gel de Campo Pulsado , Humanos , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Infecções Estafilocócicas/microbiologia , Staphylococcus epidermidis/classificação , Staphylococcus epidermidis/genética , Staphylococcus epidermidis/isolamento & purificação , Tunísia
11.
Pathol Biol (Paris) ; 55(8-9): 361-5, 2007 Nov.
Artigo em Francês | MEDLINE | ID: mdl-17905538

RESUMO

THE AIM OF STUDY: Appreciation of the frequency, the level and the genetic support of methicillin resistance. MATERIAL AND METHODS: Seventy-three strains of coagulase negative staphylococci isolated from various specimens, from January to June 2004, were studied. The phenotypic detection was carried out by disk diffusion test using oxacillin and cefoxitin disks, by the determination of oxacillin Minimal Inhibitor Concentration (E-test), by the oxacillin screening test at a concentration of 4 mug/ml and by the search of the penicillin binding protein PBP2a using the slide latex agglutination test. The results of these methods were compared to PCR of mecA gene. RESULTS: Forty-eight strains carried mecA gene whose 30 were detected by the oxacillin disk, the cefoxitin disk, the oxacillin screening test, the slide latex agglutination test and had a MIC from 24 to 256 mug/ml. Seventeen strains were not detected by oxacillin disk but by cefoxitin disk and the slide latex agglutination test. Among these strains, 13 (76%) had oxacillin MIC from 0.5 to 1,5 mug/ml and not grew on oxacillin agar screening, while 4 (24%) had oxacillin MIC from 6 to 16 mug/ml and grew on this agar. One strain had oxacillin MIC of 0,19 mug/ml and was not detected with any phenotypic method. CONCLUSION: The determination of oxacillin MIC, the search of the PBP2a or more simply the cefoxitin disk had permitted to detect the strains mecA gene (+) with resistant and pre-resistant phenotype but not the strain with sensible phenotype (2.1%).


Assuntos
Resistência a Meticilina/genética , Staphylococcus aureus/genética , Coagulase/deficiência , Coagulase/genética , Testes de Sensibilidade Microbiana , Oxacilina/farmacologia , Fenótipo , Reação em Cadeia da Polimerase , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/enzimologia
12.
Clin Microbiol Infect ; 13(8): 794-800, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17610599

RESUMO

Following systematic screening for ceftazidime-resistant (CAZ-R) Pseudomonas aeruginosa, 24 isolates producing extended-spectrum beta-lactamase (ESBL) were recovered during a 24-month period at the National Bone Marrow Transplant Centre of Tunisia. These isolates were from seven immunocompromised patients and from environmental swabs. ESBLs inhibited by clavulanic acid were detected by double-disk diffusion tests. Isoelectric focusing revealed that these isolates produced two to four beta-lactamases with pIs of 5.5, 6.1, 6.4, 7.6 or 8.2, and PCR detected the presence of bla(OXA-18), bla(SHV) and bla(TEM) genes in 24, 21 and two isolates, respectively. Pulsed-field gel electrophoresis defined two dominant genotypic groups: group A (16 isolates) and group B (four isolates). Sequencing of PCR products from representative isolates identified the bla(OXA-18) gene and revealed nucleotide sequences belonging to the bla(SHV-1) and bla(TEM-1) genes. Isolates producing OXA-18 belonged to genomic group A and were isolated from four immunocompromised patients in the haematology and graft units, and from two wash-basins in the graft unit. No immunocompromised patient harboured the clonal epidemic strain upon admission. This is the first report of the OXA-18-type ESBL in P. aeruginosa in Tunisia, and the first description of an outbreak caused by an OXA-18-producing strain of P. aeruginosa.


Assuntos
Infecção Hospitalar/microbiologia , Surtos de Doenças , Hospedeiro Imunocomprometido , Infecções por Pseudomonas/genética , Pseudomonas aeruginosa , beta-Lactamases/metabolismo , Antibacterianos/farmacologia , Transplante de Medula Óssea/efeitos adversos , Ceftazidima/farmacologia , Humanos , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/genética , Tunísia/epidemiologia , beta-Lactamases/genética
13.
Pathol Biol (Paris) ; 55(5): 230-4, 2007 Jun.
Artigo em Francês | MEDLINE | ID: mdl-17572242

RESUMO

Thirteen patients treated with gut decontamination were screened over a period of three years for digestive colonization and acquisition of resistant strains, in fact 297 strains had been isolated from 226 stool cultures within 120 enterobacteria and other species. Our study pointed out a betaLSE digestive colonization rate of 30.8% of the total enterobacteria isolated, these strains exhibit resistance of most beta-lactams especially against third generation cephalosporins. This analysis showed that these strains are endogen and specific for each patient, the common multiresistance resulted from the selective pressure of gut decontamination.


Assuntos
Enterobacteriaceae/isolamento & purificação , Neutropenia/epidemiologia , Neutropenia/microbiologia , Adulto , Enterobacteriaceae/crescimento & desenvolvimento , Enterococcus faecalis/isolamento & purificação , Escherichia coli/isolamento & purificação , Feminino , França/epidemiologia , Humanos , Masculino , Pseudomonas aeruginosa/isolamento & purificação , Staphylococcus aureus/isolamento & purificação
14.
Pathol Biol (Paris) ; 55(6): 277-82, 2007 Jul.
Artigo em Francês | MEDLINE | ID: mdl-17532578

RESUMO

Slime production, principal virulence factor of Staphylococcus epidermidis associated with catheter-related infections is mediated by icaADBC operon wich expression is subject to phase variation. Reversible transposition of IS256 element into this operon is one of the most important mechanisms of biofilm phenotypic variation. Our study compared 28 S. epidermidis strains from catheter-related infection to 28 strains from nasal carriage concerning slime production on Congo red agar plate and ica genes and IS256 presence by PCR. ica operon was present among all slime-producing strains, and was absent among slime-negative strains. Only 79% of ica-positive strains were slime producers and no insertion of IS256 element was detected inside ica genes. A significative difference was found between catheter-related infections strains and commensal ones in terms of oxacillin (67,8 versus 35,7%) and ofloxacin resistance (75 versus 35,7%), slime production (64,2 versus 28,5%), phase variability (46,4 versus 7,1%) and ica genes presence (82,1 versus 35,7%). Our study demonstrates the role of ica genes, of phenotypic variability of slime production and antibiotic multiresistance as virulence factors of S. epidermidis associated with catheter-related infections; it confirms also the complexity and the diversity of regulation mechanisms implicated in biofilm formation.


Assuntos
Cateteres de Demora/efeitos adversos , Óperon , Infecções Estafilocócicas/microbiologia , Staphylococcus epidermidis/genética , Staphylococcus epidermidis/isolamento & purificação , Antibacterianos/farmacologia , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Elementos de DNA Transponíveis , Humanos , Testes de Sensibilidade Microbiana , Infecções Estafilocócicas/etiologia , Staphylococcus epidermidis/efeitos dos fármacos
15.
Clin Microbiol Infect ; 13(1): 103-6, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17184298

RESUMO

The prevalence of macrolide-lincosamide-streptogramin (MLS) resistance phenotypes was determined among erythromycin-resistant Staphylococcus epidermidis isolates collected at the Bone Marrow Transplant Centre, Tunisia during 2002. The erm(A), erm(B), erm(C), msrA, mefA and icaA genes were detected by PCR. The vga, vgb and vat genes were amplified from pristinamycin-resistant isolates. The icaA gene was detected in 76.5% of 34 isolates examined in detail. The erm(C) (53%) and erm(A) (32%) genes predominated because of clonal dissemination, followed by msrA (15%). Gene distribution was related to the methicillin resistance pattern. The vga gene was present in combination with erm(A) in three isolates.


Assuntos
Farmacorresistência Bacteriana/genética , Neutropenia/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus epidermidis/genética , Centros Médicos Acadêmicos , Amidoidrolases/genética , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Eritromicina/farmacologia , Genes Bacterianos/genética , Humanos , Lincosamidas , Macrolídeos/farmacologia , Metiltransferases/genética , Testes de Sensibilidade Microbiana , Neutropenia/complicações , Infecções Estafilocócicas/complicações , Infecções Estafilocócicas/prevenção & controle , Staphylococcus epidermidis/efeitos dos fármacos , Estreptograminas/farmacologia , Tunísia/epidemiologia
16.
Pathol Biol (Paris) ; 54(10): 596-9, 2006 Dec.
Artigo em Francês | MEDLINE | ID: mdl-17030453

RESUMO

OBJECTIVE: Investigation of an outbreak caused by an imipenem-resistant Pseudomonas aeruginosa strain and research of its hospital reservoir. PATIENTS AND METHODS: Nine strains isolated from protected tracheal specimens during 14 weeks (October 2004 to January 2005) from 8 infants, and one strain from vacuum interrupter were studied. Epidemiological study was investigated by determination of antibiotics susceptibility, serotyping and Pulsed-field gel electrophoresis (PFGE). RESULTS: Strains were of O:12 serotype, they have the same antibiotype characterised by imipenem resistance. Strains were indistinguishable or closely related as determined by PFGE. The common source of P. aeruginosa O:12 strains was not determined, however eradication of the epidemic strain was obtained by amelioration of hygiene conditions and the change of disinfectors. CONCLUSION: Outbreak of respiratory infections due to an imipenem-resistant P. aeruginosa O:12. The common source of the epidemic strain was not determined.


Assuntos
Infecção Hospitalar/epidemiologia , Imipenem/farmacologia , Unidades de Terapia Intensiva Neonatal/estatística & dados numéricos , Infecções por Pseudomonas/epidemiologia , Pseudomonas aeruginosa , Antibacterianos/farmacologia , Infecção Hospitalar/microbiologia , Farmacorresistência Bacteriana , Humanos , Recém-Nascido , Infecções por Pseudomonas/transmissão , Pseudomonas aeruginosa/efeitos dos fármacos , Tunísia
17.
Bull Soc Pathol Exot ; 99(3): 155-9, 2006 Jul.
Artigo em Francês | MEDLINE | ID: mdl-16983816

RESUMO

In this article a Cytomegalovirus (CMV) antigenemia and semiquantitative PCR retrospective evaluation of 26 bone marrow allo-grafted patients for different haematological disease is reported. Eighteen patients had a CMV reactivation despite a prophylactic treatment, seven of those patients had both positive antigenemia pp65 and positive semi-quantitative CMV PCR. During CMV reactivation, 3 patients developed a CMV disease despite a pre-emptive therapy. The follow up of the antigenemia was performed since D21 until D100 post transplantation, the antigenemia positivity occurred at D53 and was preceded about 7 days by CMV PCR positivity The CMV disease wasn't associated with a high viral load. All patients that had CMV reactivation had a positive CMV serology before the graft, whereas only 37.5% of the patients who did not reactivate had a positive CMV serology. Respectively half patients who reactivated and only 12.5% of those who didn't had a Graft versus host disease (GVHD), witch preceded the reactivation about 21 days in six of the formers. Clinical and biological signs presented by our patients in this cases report, seems to be associated more with the GVHD than with CMV reactivation.


Assuntos
Antígenos Virais/sangue , Transplante de Medula Óssea/efeitos adversos , Infecções por Citomegalovirus/sangue , Infecções por Citomegalovirus/etiologia , Citomegalovirus/imunologia , Fosfoproteínas/sangue , Reação em Cadeia da Polimerase , Proteínas da Matriz Viral/sangue , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Reação em Cadeia da Polimerase/métodos , Prognóstico , Estudos Retrospectivos
18.
Bone Marrow Transplant ; 37(6): 595-9, 2006 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-16462754

RESUMO

In patients with central venous catheters (CVCs), catheter-related bloodstream infections (CRBI) are a prominent cause of morbidity, excess hospital costs, and in some cases mortality. The aim of this prospective study was to assess the validity of the Gram stain-acridine orange leukocyte cytospin (AOLC) test for the diagnosis of CRBI in hematopoietic stem cell transplant (HSCT) recipients with nontunnelled CVCs, using the differential-time-to-positivity (DTP)/clinical criteria as the criterion standard to define CRBIs. CVCs were externalized, nontunnelled, polyurethane double lumen catheters (Arrows, Readings, USA). All CVCs were placed in the subclavian vein by the infraclavicular approach, in the operating room. Catheters were inserted percutaneously, using the Seldinger technique. Study catheters were not exchanged over guidewires. Between May 2002 and December 2004, a total of 245 consecutive patients were included. Twenty-six of the 245 patients (10.6%) had CRBI as determined by the DTP method. The Gram stain-AOLC was positive in only two patients (7.6%) with a CRBI. Our results suggest that the Gram stain-AOLC test is not useful for the diagnosis of catheter-related bloodstream infection in HSCT recipients.2006.


Assuntos
Infecções Bacterianas/epidemiologia , Cateterismo Venoso Central/efeitos adversos , Transplante de Células-Tronco/efeitos adversos , Adolescente , Adulto , Bactérias/classificação , Bactérias/isolamento & purificação , Infecções Bacterianas/sangue , Infecções Bacterianas/diagnóstico , Criança , Pré-Escolar , Feminino , Violeta Genciana , Humanos , Masculino , Pessoa de Meia-Idade , Fenazinas , Estudos Prospectivos , Reprodutibilidade dos Testes , Transplante de Células-Tronco/métodos , Tunísia
19.
Med Mal Infect ; 35(11): 530-5, 2005 Nov.
Artigo em Francês | MEDLINE | ID: mdl-16309870

RESUMO

OBJECTIVE: Our study dealt with antibiotic resistance and serotypes of Pseudomonas aeruginosa strains isolated from immunocompromised patients in the National Bone Marrow Transplant Center of Tunis as well as molecular typing of ceftazidime resistant strains (CAZ-R). DESIGN: We studied a total of 87 non-replicate P. aeruginosa isolates from 36 patients (84 strains) or the hospital environment (3 strains). RESULTS: Rates of antimicrobial resistance were 36% for ceftazidime, 16% for imipenem, 38% for amikacin, and 57% for ciprofloxacin. The 31 CAZ-R strains were associated with O:11 serotype in 84% of the cases. Genetically characterization of CAZ-R strains by Pulsed Field Gel Electrophoresis (PFGE) after digestion of genomic DNA with SpeI revealed 2 genotypic groups. The first was composed of strains isolated from one outpatient between November 1998 and April 1999. Resistance phenotypes of these strains varied after use of antimicrobial drugs. The second was predominant (18/31 CAZ-R strains) in both hematology and graft units and persisted from June 1998 to June 2000 among 5/8 patients. These strains had O:11 serotype in 78% of the cases. The strains of this group were not isolated on patient admission and were isolated from 2 washbasins in the graft unit in May 1999. CONCLUSION: These results suggest the spread of multidrug-resistant O:11 P. aeruginosa clone from a tap water among hospitalized patients in our center, emphasizing the need of standard control of washbasins to eradicate this reservoir.


Assuntos
Antibacterianos/farmacologia , Ceftazidima/farmacologia , Hospedeiro Imunocomprometido , Pseudomonas aeruginosa/classificação , Pseudomonas aeruginosa/efeitos dos fármacos , Resistência beta-Lactâmica , Humanos , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/isolamento & purificação , Sorotipagem
20.
Bone Marrow Transplant ; 35(4): 397-401, 2005 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-15640824

RESUMO

Catheter-related bloodstream infections are associated with recognized morbidity and mortality. Accurate diagnosis of such infections results in proper management of patients and in reducing unnecessary removal of catheters. We carried out a prospective study in a bone marrow transplant unit to assess the validity of a test based on the earlier positivity of central venous blood cultures in comparison with peripheral blood cultures for predicting catheter-related bacteremia. Between May 2002 and June 2004, 38 bloodstream infections with positive simultaneous central venous catheter and peripheral vein blood cultures were included. A total of 22 patients had catheter-related bacteremias and 16 had noncatheter-related bacteremias, using the catheter-tip culture/clinical criteria as the criterion standard to define catheter-related bacteremia. Differential time to positivity of 120 min or more was associated with 86% sensitivity and 87% specificity. In conclusion, differential time to positivity of 120 min or more is sensitive and specific for catheter-related bacteremia in hematopoietic stem cell transplant recipients who have nontunnelled short-term catheters.


Assuntos
Bacteriemia/microbiologia , Cateterismo , Transplante de Células-Tronco Hematopoéticas , Adolescente , Adulto , Idoso , Bacteriemia/diagnóstico , Bacteriemia/mortalidade , Criança , Pré-Escolar , Contagem de Colônia Microbiana , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Sensibilidade e Especificidade
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