RESUMO
INTRODUCTION & AIMS: Cryopreservation of serum samples is a standard procedure for biomedical research in tertiary centers. However, studies evaluating the long-term biological stability of direct liver fibrosis markers using cryopreserved samples are scarce. METHODS: We compared the stability of hyaluronic acid (HA), tissue inhibitor of metalloproteinases (TIMP-1) and amino-terminal propeptide of type III procollagen (PIIINP) in 225 frozen serum samples of HCV-infected patients with a paired liver biopsy for up to 25 years (1990-2014). Moreover, we assessed the diagnostic accuracy (AUROC) of the Enhanced Liver Fibrosis (ELF®) score to identify significant fibrosis (F2-4) and its predictive capacity to identify clinical events during follow-up. RESULTS: Seventy-six patients (39,8%) had mild fibrosis (F0-1) and 115 (60,2%) significant fibrosis (F2-4). HA, PIIINP and TIMP-1 values remained stable during the period from 1995 to 2014 while those of 1990-94 were slightly higher. We did not find significant differences in the median ELF® values during the 20-year period from 1995-2014 in patients with mild (from 8,4 to 8,7) and significant fibrosis (from 9,9 to 10,9) (p = ns between periods and fibrosis stages). The AUROCs of ELF® to identify significant fibrosis were high in all the periods (from 0,85 to 0,91). The ELF® score showed a good predictive capability to identify clinical events during follow-up. CONCLUSIONS: The biological stability of direct serum markers (HA, PIIINP and TIMP-1) using HCV-infected samples cryopreserved for 20 years is good. Therefore, the diagnostic accuracy of the ELF® score to identify significant fibrosis and clinical events during follow-up is very high.
Assuntos
Hepatite C Crônica/complicações , Ácido Hialurônico/sangue , Cirrose Hepática/diagnóstico , Fragmentos de Peptídeos/sangue , Pró-Colágeno/sangue , Inibidor Tecidual de Metaloproteinase-1/sangue , Adulto , Idoso , Biomarcadores/sangue , Criopreservação , Feminino , Hepatite C Crônica/diagnóstico , Humanos , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Adulto JovemRESUMO
Sonication improved the diagnosis of orthopedic implant-associated infections (OIAI). We investigated the diagnostic performance of sonication fluid inoculated into blood culture bottles in comparison with that of intraoperative tissue and sonication fluid cultures. Consecutive patients with removed orthopedic hardware were prospectively included and classified as having OIAI or aseptic failure (AF) according to standardized criteria. The diagnostic procedure included the collection of five intraoperative tissue cultures and sonication of the removed device, followed by conventional culture of the sonication fluid. Cultures were incubated for 7 days (aerobic) or 14 days (anaerobic). In addition, 10 ml of sonication fluid was inoculated into each aerobic and anaerobic BacT/Alert FAN blood culture bottle and incubated in the automated blood culture system for 5 days. Of 75 included patients, 39 had OIAI and 36 AF. The sensitivity of sonication fluid inoculated into blood culture bottles (100%) was higher than that of conventional sonication fluid (87%; P = 0.05) or intraoperative tissue cultures (59%; P < 0.01). Previous antibiotic therapy reduced the culture sensitivity of conventional sonication fluid to 77% and that of intraoperative tissue to 55%, while it remained 100% for blood culture-inoculated sonication fluid. The time to positivity was shorter in blood culture-inoculated sonication fluid, with detection of 72% of microorganisms after 1 day of incubation, than for intraoperative tissue and conventional sonication fluid cultures, with detection of 18% and 28% of microorganisms, respectively. In conclusion, compared to conventional sonication fluid and intraoperative tissue cultures, sonication fluid inoculated into blood culture bottles improved the diagnosis of OIAI and considerably reduced the time to culture positivity.
Assuntos
Técnicas Microbiológicas/métodos , Próteses e Implantes/microbiologia , Infecções Relacionadas à Prótese/diagnóstico , Sonicação , Manejo de Espécimes/métodos , Adulto , Aerobiose , Idoso , Idoso de 80 Anos ou mais , Anaerobiose , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Procedimentos Ortopédicos/efeitos adversos , Sensibilidade e Especificidade , Fatores de TempoRESUMO
OBJECTIVES: The sensitivity of periprosthetic tissue culture is inadequate for the diagnosis of prosthetic joint infection (PJI). We investigated and compared the values of sonication fluid culture and periprosthetic tissue culture for diagnosing PJI. METHODS: Included were patients whose joint prosthesis had been removed for any reason. The resulting sonication fluid and periprosthetic tissues were cultured for 14 days. RESULTS: Of 231 explanted prostheses, aseptic failure was diagnosed in 162 cases (70%) and PJI in 69 (30%). In PJI cases, sonication fluid culture detected 62 microorganisms and periprosthetic tissue culture detected 45. Tissue and sonication fluid cultures showed sensitivities of 61% and 81%, respectively (p < 0.01), with specificity of 100% and 99%, respectively. On day 1, tissue and sonication fluid cultures were positive in 13% and 28% (p = 0.013) of PJI cases respectively, and on day 2, in 26% and 48% (p = 0.002) of cases. Four anaerobes grew in sonication fluid culture after 7-13 days incubation, whereas tissue culture missed 3 of these. Prolonged incubation of sonication fluid did not detect any organisms in the cases of aseptic failure. CONCLUSIONS: Sonication fluid culture provides a more rapid diagnosis and detects about 30% more pathogens, although anaerobic organisms require up to 2 weeks of incubation.
Assuntos
Artrite/diagnóstico , Técnicas Microbiológicas/métodos , Próteses e Implantes/microbiologia , Infecções Relacionadas à Prótese/diagnóstico , Sonicação/métodos , Manejo de Espécimes/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Coortes , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Sensibilidade e Especificidade , Fatores de TempoRESUMO
OBJECTIVES: Ethambutol resistance has mostly been related to mutations in the embB gene. The objective of the present study was to characterize the embB gene in a collection of ethambutol-resistant and ethambutol-susceptible isolates of Mycobacterium tuberculosis complex (MTBC) from Barcelona, and to develop a DNA microarray for the rapid detection of embB mutations in our area. METHODS: Fifty-three ethambutol-resistant and 702 ethambutol-susceptible isolates of MTBC were sequenced in internal 982-1495 bp fragments of the embB gene. In addition, a low-cost, low-density array was designed to include the embB codons identified as being most frequently mutated in our area (LD-EMB array). RESULTS: The global prevalence of embB mutations found among the ethambutol-resistant isolates was 77.4% (41/53). Substitutions in embB306 were the most common [53.7% (22/41)], followed by substitutions in embB406 [26.8% (11/41)]. The presence of mutations in embB406 was related to higher levels of ethambutol resistance and to multidrug resistance. Among unrelated isolates (from 24-locus MIRU-VNTR genotyping), the percentage of embB-mutated isolates was 72.9% (27/37)--59.3% (16/27) in embB306 and 25.9% (7/27) in embB406. None of the ethambutol-susceptible isolates studied showed a mutation in codon 306 or 406. The LD-EMB array showed 100% sensitivity and specificity in identifying the main embB substitutions in our area. CONCLUSIONS: Mutations at codons 306 and 406 of embB have a relevant role in resistance to ethambutol in our area. The LD-EMB array developed in this study would appear to be a good molecular test for rapid detection of ethambutol resistance.
Assuntos
Antituberculosos/farmacologia , Farmacorresistência Bacteriana , Etambutol/farmacologia , Mutação de Sentido Incorreto , Mycobacterium tuberculosis/genética , Pentosiltransferases/genética , Códon , Genótipo , Humanos , Mycobacterium tuberculosis/isolamento & purificação , Análise de Sequência com Séries de Oligonucleotídeos , Análise de Sequência de DNA , EspanhaRESUMO
BACKGROUND: The outcome of revision surgery depends on accurate determination of the cause of prosthesis failure because treatment differs profoundly among aseptic loosening, mechanical failure, and prosthetic joint infections (PJI). QUESTIONS/PURPOSES: We sought to determine (1) the predictive role of the interval from primary to revision surgery in determining the reason for prosthesis failure of a hip, knee, shoulder, or elbow arthroplasty, and (2) whether positive cultures during revision surgery for aseptic loosening were associated with shorter event-free survival of the prosthesis. METHODS: All patients undergoing revision surgery between July 2010 and January 2012 were included in a prospective cohort of 112 patients, and were classified as having had failure from aseptic loosening (56%), mechanical failure (15%), or PJI (29%). To make the diagnosis of PJI, at surgery we used a standardized enhanced diagnostic approach in all patients including sampling of five periprosthetic tissue specimens, sonication of removed prosthetic components, prolonged incubation of aerobic and anaerobic cultures, and multiplex PCR of sonication fluid in aseptic loosening cases. Kaplan-Meier survival and Cox proportional hazards regression analysis were performed. RESULTS: The median time from primary to revision surgery was (p < 0.001) longer for patients with aseptic loosening (7.8 years) than for patients with mechanical failure (1.6 years) or PJI (2 years). No difference in the time to revision was observed for patients with aseptic loosening with positive or negative microbiological cultures (p = 0.594). Propionibacterium acnes was cultured below the established microbiological criteria for positivity in 12 (19%) procedures that had been presumed to have been revisions for aseptic loosening. CONCLUSIONS: PJI should be considered in all revisions performed within 2 years of implantation even in the absence of clinical or laboratory findings suggestive for infection. However, the growth of low-virulence microorganisms below the cut-off in revisions for apparent aseptic loosening is not associated with early prosthesis failure.
Assuntos
Artroplastia de Substituição/efeitos adversos , Prótese Articular/efeitos adversos , Falha de Prótese , Infecções Relacionadas à Prótese/etiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Artroplastia de Substituição/instrumentação , Distribuição de Qui-Quadrado , Feminino , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais , Estudos Prospectivos , Infecções Relacionadas à Prótese/diagnóstico , Infecções Relacionadas à Prótese/microbiologia , Infecções Relacionadas à Prótese/cirurgia , Reoperação , Fatores de Risco , Espanha , Estresse Mecânico , Fatores de Tempo , Adulto JovemRESUMO
In cases of multidrug-resistant tuberculosis, it is crucial to rule out resistance to second-line antituberculous (anti-TB) agents. In the present study, a low-cost low-density DNA array including four genetic regions (rrs 530 loop, rrs 1400, rpsL and gyrA) was designed for the rapid detection of the most important mutations related to anti-TB injectable drugs (mainly streptomycin) and fluoroquinolone resistance (LD-SQ array). A total of 108 streptomycin- and/or ofloxacin-resistant and 20 streptomycin- and ofloxacin-susceptible Mycobacterium tuberculosis clinical isolates were analysed with the array. The results obtained were compared with sequencing data and phenotypic susceptibility pattern. The LD-SQ array offered a good sensitivity compared to sequencing, especially among resistant strains: 92.5% (37/40) for streptomycin and 87.5% (7/8) for fluoroquinolones. Therefore, this array could be considered a good approach for the rapid detection of mutations related to streptomycin and fluoroquinolone resistance. On the other hand, there were discordant results in 16 resistant strains and six susceptible isolates, mostly concerning the gyrA region, in which the existence of polymorphisms next to informative positions might cause cross-hybridization. These discrepancies were caused by some technical limitations; consequently, the present array should be considered as a first-step prior to a forthcoming optimized version of the array.
Assuntos
Antituberculosos/farmacologia , Fluoroquinolonas/farmacologia , Mycobacterium tuberculosis/efeitos dos fármacos , Estreptomicina/farmacologia , Tuberculose Resistente a Múltiplos Medicamentos/diagnóstico , Análise Mutacional de DNA/métodos , DNA Bacteriano/genética , Farmacorresistência Bacteriana Múltipla/genética , Humanos , Testes de Sensibilidade Microbiana/métodos , Mutação , Mycobacterium tuberculosis/genética , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Tuberculose Resistente a Múltiplos Medicamentos/microbiologiaRESUMO
Biofilm removal efficacy of vortexing alone was compared with the standard vortexing-sonication procedure. Among 135 removed prostheses, 35 were diagnosed with infection and 100 with aseptic failure. At a cutoff of ≥ 50 CFU/ml, sonication was more sensitive than vortexing (60% versus 40%, P = 0.151), while the specificity was 99% for both methods.
Assuntos
Artrite Infecciosa/diagnóstico , Artroplastia de Substituição/efeitos adversos , Infecções Relacionadas à Prótese/diagnóstico , Sonicação , Adulto , Idoso , Idoso de 80 Anos ou mais , Artrite Infecciosa/microbiologia , Carga Bacteriana/métodos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infecções Relacionadas à Prótese/microbiologiaRESUMO
OBJECTIVE: Cultures have limited sensitivity in the diagnosis of prosthetic joint infection (PJI), especially in low-grade infections. We assessed the value of multiplex PCR in differentiating PJI from aseptic failure (AF). METHODS: Included were patients in whom the joint prosthesis was removed and submitted for sonication. The resulting sonication fluid was cultured and investigated by multiplex PCR, and compared with periprosthetic tissue culture. RESULTS: Among 86 explanted prostheses (56 knee, 25 hip, 3 elbow and 2 shoulder prostheses), AF was diagnosed in 62 cases (72%) and PJI in 24 cases (28%). PJI was more common detected by multiplex PCR (n=23, 96%) than by periprosthetic tissue (n=17, 71%, p=0.031) or sonication fluid culture (n=16, 67%, p=0.016). Among 12 patients with PJI who previously received antibiotics, periprosthetic tissue cultures were positive in 8 cases (67%), sonication fluid cultures in 6 cases (50%) and multiplex PCR in 11 cases (92%). In AF cases, periprosthetic tissue grew organisms in 11% and sonication fluid in 10%, whereas multiplex PCR detected no organisms. CONCLUSIONS: Multiplex PCR of sonication fluid demonstrated high sensitivity (96%) and specificity (100%) for diagnosing PJI, providing good discriminative power towards AF, especially in patients previously receiving antibiotics.
Assuntos
Infecções Bacterianas/diagnóstico , Prótese Articular/microbiologia , Reação em Cadeia da Polimerase Multiplex/métodos , Infecções Relacionadas à Prótese/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Bactérias/genética , Bactérias/isolamento & purificação , Infecções Bacterianas/microbiologia , Distribuição de Qui-Quadrado , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Infecções Relacionadas à Prótese/microbiologia , Sensibilidade e Especificidade , SonicaçãoRESUMO
BACKGROUND: We aimed to assess the hospital economic costs of nosocomial multi-drug resistant Pseudomonas aeruginosa acquisition. METHODS: A retrospective study of all hospital admissions between January 1, 2005, and December 31, 2006 was carried out in a 420-bed, urban, tertiary-care teaching hospital in Barcelona (Spain). All patients with a first positive clinical culture for P. aeruginosa more than 48 h after admission were included. Patient and hospitalization characteristics were collected from hospital and microbiology laboratory computerized records. According to antibiotic susceptibility, isolates were classified as non-resistant, resistant and multi-drug resistant. Cost estimation was based on a full-costing cost accounting system and on the criteria of clinical Activity-Based Costing methods. Multivariate analyses were performed using generalized linear models of log-transformed costs. RESULTS: Cost estimations were available for 402 nosocomial incident P. aeruginosa positive cultures. Their distribution by antibiotic susceptibility pattern was 37.1% non-resistant, 29.6% resistant and 33.3% multi-drug resistant. The total mean economic cost per admission of patients with multi-drug resistant P. aeruginosa strains was higher than that for non-resistant strains (15,265 vs. 4,933 Euros). In multivariate analysis, resistant and multi-drug resistant strains were independently predictive of an increased hospital total cost in compared with non-resistant strains (the incremental increase in total hospital cost was more than 1.37-fold and 1.77-fold that for non-resistant strains, respectively). CONCLUSIONS: P. aeruginosa multi-drug resistance independently predicted higher hospital costs with a more than 70% increase per admission compared with non-resistant strains. Prevention of the nosocomial emergence and spread of antimicrobial resistant microorganisms is essential to limit the strong economic impact.
Assuntos
Antibacterianos/farmacologia , Infecção Hospitalar/economia , Farmacorresistência Bacteriana Múltipla , Custos Hospitalares/tendências , Infecções por Pseudomonas/economia , Pseudomonas aeruginosa/efeitos dos fármacos , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/uso terapêutico , Custos e Análise de Custo , Feminino , Hospitalização/economia , Hospitalização/tendências , Hospitais de Ensino , Humanos , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa/isolamento & purificação , Estudos Retrospectivos , EspanhaRESUMO
The authors report a case of an inguinal bubo in a young man caused by an anaerobe, Prevotella bivia, which was acquired during oral sexual intercourse. As far as the authors know, this is the first reported case of a sexually transmitted infection by Prevotella. Prevotella spp. inhabit the oral cavity and are highly prevalent in bacterial vaginosis, a polymicrobial syndrome resulting from replacement of the normal vaginal Lactobacillus spp. flora by high concentrations of anaerobic microorganisms such as Prevotella spp., Mobiluncus spp., Gardnerella vaginalis and other uncultivated anaerobes.
Assuntos
Infecções por Bacteroidaceae/transmissão , Mordeduras Humanas/complicações , Canal Inguinal , Infecções Intra-Abdominais/microbiologia , Prevotella , Comportamento Sexual , Adulto , Humanos , Masculino , Síndrome , Tomografia Computadorizada por Raios XRESUMO
OBJECTIVES: To analyze the characteristics of infection, adequacy of empirical treatment and outcome of patients with community-onset healthcare-associated (HCA) urinary tract infections (UTI) and compare them with hospital (HA) and community-acquired (CA) UTI. METHODS: Prospective observational cohort study performed at a university 600-bed hospital between July 2009 and February 2010. Patients with UTI requiring hospital admission were included. Epidemiological, clinical and outcome data were recorded. RESULTS: 251 patients were included. Patients with community-onset HCA UTI were older, had more co-morbidities and had received previous antimicrobial treatment more frequently than CA UTI (p = 0.02, p = 0.01 and p < 0.01). ESBL-Escherichia coli and Pseudomonas aeruginosa infections were more frequent in HCA than in CA UTI (p = 0.03 and p < 0.01). Inadequate empirical treatment was not significantly different between community-onset HCA and CA. Factors related to mortality were P. aeruginosa infection (OR 6.51; 95%CI: 1.01-41.73), diabetes mellitus (OR 22.66; 95%CI: 3.61-142.21), solid neoplasia (OR 22.48; 95%CI: 3.38-149.49) and age (OR 1.15; 95%CI 1.03-1.28). CONCLUSIONS: Epidemiological, clinical and microbiological features suggest that community-onset HCA UTI is different from CA and similar to HA UTI. However, in our series inadequate empirical antimicrobial therapy and mortality were not significantly higher in community-onset HCA than in CA UTI.
Assuntos
Infecções Bacterianas/epidemiologia , Infecções Comunitárias Adquiridas/epidemiologia , Infecção Hospitalar/epidemiologia , Infecções Urinárias/epidemiologia , Adulto , Distribuição por Idade , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/uso terapêutico , Bactérias/classificação , Bactérias/isolamento & purificação , Infecções Bacterianas/tratamento farmacológico , Infecções Bacterianas/microbiologia , Infecções Bacterianas/patologia , Estudos de Coortes , Infecções Comunitárias Adquiridas/tratamento farmacológico , Infecções Comunitárias Adquiridas/microbiologia , Infecções Comunitárias Adquiridas/patologia , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/microbiologia , Infecção Hospitalar/patologia , Farmacorresistência Bacteriana , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Análise de Sobrevida , Resultado do Tratamento , Infecções Urinárias/tratamento farmacológico , Infecções Urinárias/microbiologia , Infecções Urinárias/patologiaRESUMO
OBJECTIVES: We analysed the ability of Mycobacterium tuberculosis clinical isolates to penetrate and grow inside murine macrophages as a surrogate of fitness. METHODS: Thirty-five drug-resistant and 10 drug-susceptible M. tuberculosis isolates were studied in a murine macrophage model from the J774.2 cell line in a 6 day protocol, performing semi-quantitative counts in Middlebrook 7H11 medium. The mycobacterial penetration index (MPI) after infection and the mycobacterial growth ratio (MGR) inside the macrophages were determined to evaluate the fitness of isolates. RESULTS: Isolates with the katG S315T mutation and multidrug-resistant (MDR) isolates had a significantly lower MGR compared with drug-susceptible isolates. The MPI of the isolates with the katG S315T mutation showed a significant decrease compared with the MPI of those without this mutation. A trend to significantly lower values was also observed on comparing the MPI of the MDR isolates with that of the drug-susceptible isolates and the isolates resistant to isoniazid. CONCLUSIONS: The isoniazid-resistant and MDR isolates with mutations in the katG gene showed decreased multiplication inside murine macrophages, suggesting a lower fitness of M. tuberculosis with these resistance patterns.
Assuntos
Antituberculosos/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Macrófagos/microbiologia , Mycobacterium tuberculosis/crescimento & desenvolvimento , Animais , Proteínas de Bactérias/genética , Catalase/genética , Linhagem Celular , RNA Polimerases Dirigidas por DNA , Humanos , Isoniazida/farmacologia , Camundongos , Testes de Sensibilidade Microbiana , Mutação , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/isolamento & purificação , Oxirredutases/genéticaRESUMO
OBJECTIVES: To determine the proportion and type of mutations in Mycobacterium tuberculosis isolates resistant to streptomycin, and their relationship with the level of resistance and with the epidemiological molecular pattern of the isolates. METHODS: Sixty-nine streptomycin-resistant isolates from a M. tuberculosis strain collection (1995-2005) from Barcelona were studied. The MIC of streptomycin for each isolate was determined using the proportions method with Middlebrook 7H11 medium. The entire rpsL gene and two specific fragments of the rrs gene (the 530 loop and the 912 region) were sequenced. IS6110-restriction fragment length polymorphism and spoligotyping were performed in each isolate. RESULTS: Twenty-six (26/69, 37.7%) streptomycin-resistant isolates presented a mutation in either the rpsL gene and/or the rrs530 loop, with no mutation in the rrs912 region. Seventeen (24.6%) isolates showed rpsL mutations (codons 43 and 88) associated with high MIC levels. Nine (13.0%) isolates had alterations in the rrs gene (A513T, A513C and C516T). Nineteen isolates (19/64, 29.7%) were classified into seven clusters (containing 2-5 isolates per cluster). Nineteen different spoligotype patterns were found. All the LAM3 spoligotype isolates (10/67, 14.9%) were associated with a C491T change in the rrs gene, being also observed in all LAM3 streptomycin-susceptible isolates. CONCLUSIONS: Mutations in the rpsL and rrs genes were detected in 37.7% of streptomycin-resistant M. tuberculosis isolates. High-level resistance was associated with mutations in the rpsL gene, whereas wild-type isolates showed low MIC levels. The presence of the C491T substitution in the rrs gene in streptomycin-susceptible and -resistant isolates demonstrates that this change is an epidemiological marker associated with LAM3 sublineage.
Assuntos
Antituberculosos/farmacologia , Farmacorresistência Bacteriana , Mutação de Sentido Incorreto , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/genética , Estreptomicina/farmacologia , Tuberculose/microbiologia , Técnicas de Tipagem Bacteriana , Impressões Digitais de DNA , Elementos de DNA Transponíveis , DNA Bacteriano/genética , Humanos , Testes de Sensibilidade Microbiana , Mycobacterium tuberculosis/isolamento & purificação , Proteínas Ribossômicas/genética , EspanhaRESUMO
OBJECTIVES: We describe 12 VIM-1-producing strains (7 Enterobacter cloacae, 2 Klebsiella pneumoniae and 3 clonal Klebsiella oxytoca strains) detected among clinically relevant Enterobacteriaceae isolates from routine cultures at the Hospital del Mar (Barcelona, Spain) from December 2006 to May 2007. METHODS: Susceptibility to carbapenems was evaluated with the MicroScan system. beta-Lactamases were identified by PCR and sequencing. Clonal relationships between the isolates were analysed by PFGE. Transferability of the enzymes was tested by conjugation. Plasmid characterization was performed by PCR-based replicon typing and PFGE with S1 nuclease digestion of whole genomic DNA. The PFGE gels were then transferred and hybridized. RESULTS: The disc diffusion method correctly identified five of the seven E. cloacae isolates as intermediate or resistant strains. All isolates produced the VIM-1 enzyme. Three E. cloacae and three K. oxytoca strains were also CTX-M-9-producing strains, and one E. cloacae was also a CTX-M-3-producing strain. The plasmids carrying the bla(VIM) gene, of unknown incompatibility group, had a size of approximately 75 kb (eight strains) or 40 kb (three strains) and also contained the qnrS and the aac(6')-Ib-cr genes. In the remaining strain the bla(VIM-1) gene was found in an HI2 plasmid of 290 kb together with bla(CTX-M-9), qnrA, qnrS and the aac(6')-Ib-cr genes. CONCLUSIONS: The results showed a linkage between the bla(VIM-1) and the qnrS and the aac(6')-Ib-cr genes, and between the bla(CTX-M-9) and the qnrA genes.
Assuntos
Proteínas de Bactérias/genética , Enterobacter cloacae/genética , Infecções por Enterobacteriaceae/microbiologia , Klebsiella oxytoca/genética , Klebsiella pneumoniae/genética , Plasmídeos , beta-Lactamases/genética , Idoso , Idoso de 80 Anos ou mais , Técnicas de Tipagem Bacteriana , Análise por Conglomerados , Conjugação Genética , Impressões Digitais de DNA , DNA Bacteriano/química , DNA Bacteriano/genética , Farmacorresistência Bacteriana , Eletroforese em Gel de Campo Pulsado , Enterobacter cloacae/isolamento & purificação , Feminino , Hospitais , Humanos , Klebsiella oxytoca/isolamento & purificação , Klebsiella pneumoniae/isolamento & purificação , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , EspanhaRESUMO
BACKGROUND: Recent reports have suggested that anastomotic leakage is associated with greater rates of tumor recurrence and cancer-specific mortality after surgery for colorectal cancer. The impact of postoperative intra-abdominal infection on long-term oncologic results, however, is still controversial, and no direct causal relationship has been found between both processes. Our aim was to investigate the influence of postoperative intraabdominal infection on angiogenesis and tumor growth in an animal model of colon cancer. METHODS: Balb/c mice were randomized immediately after injection of 5x10(6) B51LiM cells into the cecal wall into 2 groups: cecal resection without postoperative infection (group 1), and cecal resection with postoperative intra-abdominal infection (group 2). A total of 18 days after cell injection, cecectomy was performed, and infection was induced in group 2 by intraperitoneal injection of 3x10(8) colony-forming units of Bacteroides fragilis. On postoperative day 12, the mice were killed. RESULTS: Comparing group 1 with group 2, tumor recurrence was more frequent in animals with intraabdominal infection (65% vs 100%, respectively; P=02). VEGF serum levels were greater at the time of sacrifice in the group with infection (11+/-10 vs 30+/-23 pg/mL; P<.05). Tumor angiogenesis was also increased in the postoperative infection group. The mean (+/-standard deviation) microvessel density was 16+/-7 versus 28+/-11 vessels per high-power field (P<.05). CONCLUSION: We concluded that postoperative intra-abdominal infection increases angiogenesis and tumor recurrence after operative excision of a colon cancer in mice.
Assuntos
Adenocarcinoma/cirurgia , Neoplasias do Colo/cirurgia , Recidiva Local de Neoplasia/etiologia , Neovascularização Patológica/etiologia , Infecção da Ferida Cirúrgica/complicações , Adenocarcinoma/patologia , Animais , Linhagem Celular Tumoral , Colo/patologia , Neoplasias do Colo/patologia , Camundongos , Camundongos Endogâmicos BALB C , Peritonite/complicaçõesRESUMO
OBJECTIVES: To determine whether the incidence of bacteremia after transrectal ultrasound-guided prostate biopsy (TRUSGPB) significantly diminishes with the setting up of a new preventive protocol. This protocol was set up after detecting an augmented incidence of bacteremia after TRUSGPB with a high prevalence of antibiotic-resistant microorganisms. METHODS: Retrospective descriptive and prospective intervention study performed at a University Hospital. PARTICIPANTS: Patients undergoing TRUSGPB under the old preventive protocol (January 2006-February 2007), that is, amoxicillin-clavulanate 500 mg tid the day before, the day of the procedure, and 1 day after the procedure, and after setting up a new protocol (March 2007-April 2008), that is, 2 g cefoxitin 1 hour before the procedure and ciprofloxacin 750 mg p.o. bid the day before, the day of the procedure, and 3 days after the procedure; dipstick urinalysis was performed before the procedure, and patients with positive results were not biopsied. RESULTS: Incidence of bacteremia with old and new protocols: 9 of 204 procedures (4.4%) vs 2 of 207 (0.9%), (P = .03). Four isolates (44.4%) under the old protocol produced extended-spectrum beta-lactamase (ESBL). With the new protocol, 2 (0.9%) cases of non-ESBL Escherichia coli bacteremia were observed. Sixty-five (23.8%) cases were not biopsied because of positive result of dipstick urinalysis, lack of antibiotic prophylaxis adherence, or altered coagulation parameters. CONCLUSIONS: Antibiotic prophylaxis for TRUSGPB should take into account local resistance patterns. Cefoxitin could be used as prophylaxis in centers with high prevalence of ESBL enterobacteriaceae. Before TRUSGPB, excluding patients with positive results of dipstick urinalysis is an advisable practice.
Assuntos
Bacteriemia/etiologia , Bacteriemia/prevenção & controle , Infecções por Enterobacteriaceae/etiologia , Infecções por Enterobacteriaceae/prevenção & controle , Enterobacteriaceae/enzimologia , Próstata/diagnóstico por imagem , Próstata/patologia , beta-Lactamases/biossíntese , Idoso , Bacteriemia/epidemiologia , Bacteriemia/microbiologia , Biópsia/efeitos adversos , Biópsia/métodos , Protocolos Clínicos , Infecções por Enterobacteriaceae/epidemiologia , Humanos , Incidência , Masculino , Estudos Retrospectivos , UltrassonografiaRESUMO
The aim of this study was to analyze the factors associated with conventional contact tracing (CCT) and molecular epidemiology (ME) methods in assessing tuberculosis (TB) transmission, comparing the populations studied and the epidemiological links established by both methods. Data were obtained from TB case and CCT registries, and ME was performed using IS6110-based restriction fragment length polymorphism (RFLP) analysis and mycobacterial interspersed repetitive unit 12 (MIRU12) typing as a secondary typing method. During two years (2003 and 2004), 892 cases of TB were reported, of which 687 (77%) were confirmed by culture. RFLP analysis was performed with 463 (67.4%) of the 687 isolated strains, and MIRU12 types in 75 strains were evaluated; 280 strains (60.5%) had a unique RFLP pattern, and 183 (39.5%) shared patterns, grouping into 65 clusters. CCT of 613 (68.7%) of 892 cases detected 44 clusters involving 101 patients. The results of both CCT and ME methods yielded 96 clusters involving 255 patients. The household link was the one most frequently identified by CCT (corresponding to 80.7% of the cases clustered by this method), whereas nonhousehold and unknown links were associated with 94.1% of the strains clustered by ME. When both methods were used in 351 cases (39.3%), they showed the same results in 214 cases (61%). Of the remainder, 106 (30.2%) were clustered only by ME, 19 (5.5%) were clustered only by CCT, and 12 (3.4%) were clustered by both methods but into different clusters. Patients with factors potentially associated with social problems were less frequently studied by CCT (P = 0.002), whereas patients of <15 years of age, most with negative cultures, were less frequently studied by ME (P = 0.005). Significant differences in the populations studied by ME versus CCT were observed, possibly explaining the scarce correlation found between the results of these methods. Moreover, ME allowed the detection of nonhousehold contact relationships, whereas CCT was more useful for tracing transmission chains involving patients of <15 years of age. In conclusion, the two methods are complementary, suggesting the need to improve the methodology of contact study protocols.