RESUMO
The emergence of cell therapy programs in large academic centers has led to an increasing demand for clinical laboratories to assist with product sterility testing. Automated blood culture systems have shown promise as alternatives to the manual USP<71> compendial method, but current published data are limited by small organism test sets, particularly for molds. In 2015, failure of the Bactec FX system to detect mold contamination in two products prompted us to evaluate three test systems (compendial USP<71>, Bactec FX, and BacT/Alert Dual-T) over seven different culture combinations, using 118 challenge organisms representative of the NIH current good manufacturing practice (cGMP) environment. At <96 h and <144 h for bacterial and fungal detection, respectively, the compendial USP<71> method significantly outperformed the Bactec FX system (84.7% versus 64.4%; P = 0.0006) but not the BacT/Alert system at 32.5°C (78.8%; P = 0.3116). Extended incubation to 360 h with terminal visual inspection improved sensitivity, without a significant difference between compendial USP<71> and BacT/Alert testing (95.7% versus 89.0%; P = 0.0860); both systems were better than the Bactec FX system (71.2%; P < 0.0001 and P = 0.0003, respectively). The Bactec FX and BacT/Alert systems performed equivalently for 30 isolates derived from clinical bloodstream infections, confirming system optimization for clinical organisms rather than environmental contaminants. Paired Sabouraud dextrose agar (SDA) plates were always positive for fungi within the acceptable time frame. This study shows that the Bactec FX system is suboptimal for product sterility testing, and it provides strong data to support the use of BacT/Alert testing at 32.5°C paired with a supplemental SDA plate as an acceptable alternative to the compendial USP<71> method for product sterility testing.
Assuntos
Terapia Baseada em Transplante de Células e Tecidos/efeitos adversos , Contaminação de Medicamentos , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/prevenção & controle , Técnicas Microbiológicas/métodos , Terapia Baseada em Transplante de Células e Tecidos/métodos , Sensibilidade e EspecificidadeRESUMO
Diagnostics and research analyses involving samples containing maximum-containment viruses present unique challenges, and inactivation protocols compatible with downstream testing are needed. Our aim was to identify a validated viral inactivation protocol compatible with bacterial identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). We assessed a panel of bacteria with 6 validated maximum-containment virus-inactivation protocols and report that inactivation with TRIzol or γ-irradiation is compatible with MALDI-TOF MS. The availability, simplicity, and rapidity of TRIzol inactivation make this method the more suitable choice.
Assuntos
Bactérias/efeitos da radiação , Coinfecção/virologia , Inativação de Vírus/efeitos da radiação , Vírus/efeitos da radiação , Humanos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodosRESUMO
BACKGROUND: Klebsiella pneumoniae can be a serious pathogen in non-human primates, particularly Neotropical monkeys. METHODS: During a K. pneumoniae outbreak in an owl monkey research colony, 13 K. pneumoniae isolates were DNA fingerprinted by automated repetitive extragenic palindromic-polymerase chain reaction and the profiles compared to isolates obtained from other non-human primate species during the same time period and isolates from previous outbreaks. RESULTS: Eleven different types of K. pneumoniae were circulating in the owl monkey colony at the time of the outbreak. When comparing owl monkey isolates relatedness to previous colony outbreak isolates and squirrel monkey and capuchin monkey isolates, all were different. CONCLUSIONS: These results agree with recent reports where K. pneumoniae nosocomial isolates in hospital settings can have high genetic diversity, and multiple strains can be circulating simultaneously. This potential genetic diversity should be considered when designing strategies for controlling K. pneumoniae outbreaks in captive non-human primate colonies.
Assuntos
Aotidae , Surtos de Doenças , Variação Genética , Infecções por Klebsiella/epidemiologia , Klebsiella pneumoniae/genética , Doenças dos Macacos/epidemiologia , Animais , Animais de Laboratório , Feminino , Infecções por Klebsiella/microbiologia , Masculino , Doenças dos Macacos/microbiologiaRESUMO
Carbapenem-resistant Enterobacteriaceae (CRE) have spread globally and represent a serious and growing threat to public health. Rapid methods for tracking plasmids carrying carbapenemase genes could greatly benefit infection control efforts. Here, we demonstrate that real-time, direct tracking of a single plasmid in a bacterial strain responsible for an outbreak is possible using a commercial matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) system. In this case, we retrospectively tracked the bla(KPC) carbapenemase gene-bearing pKpQIL plasmid responsible for a CRE outbreak that occurred at the NIH Clinical Center in 2011. An â¼ 11,109-Da MS peak corresponding to a gene product of the bla(KPC) pKpQIL plasmid was identified and characterized using a combination of proteomics and molecular techniques. This plasmid peak was present in spectra from retrospectively analyzed K. pneumoniae outbreak isolates, concordant with results from whole-genome sequencing, and absent from a diverse control set of bla(KPC)-negative clinical Enterobacteriaceae isolates. Notably, the gene characterized here is located adjacent to the bla(KPC) Tn4401 transposon on the pKpQIL plasmid. Sequence analysis demonstrates the presence of this gene in other bla(KPC) Tn4401-containing plasmids and suggests that this signature MS peak may be useful in tracking other plasmids conferring carbapenem resistance. Plasmid identification using this MALDI-TOF MS method was accomplished in as little as 10 min from isolated colonies and 30 min from positive (spiked) blood cultures, demonstrating the potential clinical utility for real-time plasmid tracking in an outbreak.
Assuntos
Técnicas de Tipagem Bacteriana/métodos , Surtos de Doenças , Infecções por Enterobacteriaceae/epidemiologia , Enterobacteriaceae/classificação , Plasmídeos/análise , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Resistência beta-Lactâmica , Antibacterianos/farmacologia , Proteínas de Bactérias/análise , Proteínas de Bactérias/química , Carbapenêmicos/farmacologia , DNA Bacteriano/química , DNA Bacteriano/genética , Enterobacteriaceae/química , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Infecções por Enterobacteriaceae/microbiologia , Genes Bacterianos , Humanos , Epidemiologia Molecular/métodos , Peso Molecular , Análise de Sequência de DNA , Fatores de TempoRESUMO
Acinetobacter baumannii is an emerging human pathogen and a significant cause of nosocomial infections among hospital patients worldwide. The enormous increase in multidrug resistance among hospital isolates and the recent emergence of pan-drug-resistant strains underscores the urgency to understand how A. baumannii evolves in hospital environments. To this end, we undertook a genomic study of a polyclonal outbreak of multidrug-resistant A. baumannii at the research-based National Institutes of Health Clinical Center. Comparing the complete genome sequences of the three dominant outbreak strain types enabled us to conclude that, despite all belonging to the same epidemic lineage, the three strains diverged before their arrival at the National Institutes of Health. The simultaneous presence of three divergent strains from this lineage supports its increasing prevalence in international hospitals and suggests an ongoing adaptation to the hospital environment. Further genomic comparisons uncovered that much of the diversification that occurred since the divergence of the three outbreak strains was mediated by homologous recombination across 20% of their genomes. Inspection of recombinant regions revealed that several regions were associated with either the loss or swapping out of genes encoding proteins that are exposed to the cell surface or that synthesize cell-surface molecules. Extending our analysis to a larger set of international clinical isolates revealed a previously unappreciated ability of A. baumannii to vary surface molecules through horizontal gene transfer, with subsequent intraspecies dissemination by homologous recombination. These findings have immediate implications in surveillance, prevention, and treatment of A. baumannii infections.
Assuntos
Acinetobacter baumannii/genética , Genoma Bacteriano/genética , Recombinação Genética , Infecções por Acinetobacter/epidemiologia , Acinetobacter baumannii/classificação , Infecção Hospitalar/genética , Epidemias , Especiação Genética , Hospitais , Humanos , Dados de Sequência MolecularRESUMO
Streptococcus intermedius caused a liver abscess in a patient with chronic granulomatous disease (CGD). In contrast to typical staphylococcal abscesses in CGD, this abscess was liquid, easily drained, and resolved without surgery or steroids. This case and literature review provide insight into this organism's pathogenesis, including in CGD.
Assuntos
Doença Granulomatosa Crônica/complicações , Abscesso Hepático/microbiologia , Infecções Estreptocócicas/complicações , Adulto , Doença Granulomatosa Crônica/microbiologia , Humanos , Abscesso Hepático/complicações , Masculino , Streptococcus intermediusRESUMO
Chronic granulomatous disease (CGD) is characterized by frequent infections, most of which are curable. Granulibacter bethesdensis is an emerging pathogen in patients with CGD that causes fever and necrotizing lymphadenitis. However, unlike typical CGD organisms, this organism can cause relapse after clinical quiescence. To better define whether infections were newly acquired or recrudesced, we use comparative bacterial genomic hybridization to characterize 11 isolates obtained from 5 patients with CGD from North and Central America. Genomic typing showed that 3 patients had recurrent infection months to years after apparent clinical cure. Two patients were infected with the same strain as previously isolated, and 1 was infected with a genetically distinct strain. This organism is multidrug resistant, and therapy required surgery and combination antimicrobial drugs, including long-term ceftriaxone. G. bethesdensis causes necrotizing lymphadenitis in CGD, which may recur or relapse.
Assuntos
Acetobacteraceae , Doenças Transmissíveis Emergentes/complicações , Doenças Transmissíveis Emergentes/microbiologia , Infecções por Bactérias Gram-Negativas/complicações , Infecções por Bactérias Gram-Negativas/microbiologia , Doença Granulomatosa Crônica/complicações , Doença Granulomatosa Crônica/microbiologia , Acetobacteraceae/classificação , Acetobacteraceae/efeitos dos fármacos , Acetobacteraceae/genética , Acetobacteraceae/isolamento & purificação , Adolescente , Adulto , Sequência de Bases , Doenças Transmissíveis Emergentes/diagnóstico , Doenças Transmissíveis Emergentes/tratamento farmacológico , Primers do DNA/genética , Genoma Bacteriano , Instabilidade Genômica , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Humanos , Masculino , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , RecidivaRESUMO
The epidemiology of Burkholderia infection in persons with chronic granulomatous disease is poorly understood. We used species-specific polymerase chain reaction-based assays and genotyping analyses to identify 32 strains representing 9 Burkholderia species among 50 isolates recovered from 18 patients with chronic granulomatous disease. We found that recurrent pulmonary infection with distinct Burkholderia strains is common in chronic granulomatous disease.
Assuntos
Infecções por Burkholderia/epidemiologia , Burkholderia/isolamento & purificação , Doença Granulomatosa Crônica/complicações , Pneumonia Bacteriana/epidemiologia , Burkholderia/classificação , Burkholderia/genética , Infecções por Burkholderia/microbiologia , DNA Bacteriano/genética , Genótipo , Humanos , Pneumonia Bacteriana/microbiologia , Reação em Cadeia da Polimerase/métodos , RecidivaAssuntos
Técnicas de Tipagem Bacteriana/métodos , Surtos de Doenças , Infecções por Enterobacteriaceae/epidemiologia , Enterobacteriaceae/classificação , Plasmídeos/análise , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Resistência beta-Lactâmica , HumanosRESUMO
Mycobacterium abscessus is the most common cause of rapidly growing mycobacterial chronic lung disease. Recently, two new M. abscessus-related species, M. massiliense and M. bolletii, have been described. Health care-associated outbreaks have recently been investigated by the use of molecular identification and typing tools; however, very little is known about the natural epidemiology and pathogenicity of M. massiliense or M. bolletii outside of outbreak situations. The differentiation of these two species from M. abscessus is difficult and relies on the sequencing of one or more housekeeping genes. We performed extensive molecular identification and typing of 42 clinical isolates of M. abscessus, M. massiliense, and M. bolletii from patients monitored at the NIH between 1999 and 2007. The corresponding clinical data were also examined. Partial sequencing of rpoB, hsp65, and secA led to the unambiguous identification of 26 M. abscessus isolates, 7 M. massiliense isolates, and 2 M. bolletii isolates. The identification results for seven other isolates were ambiguous and warranted further sequencing and an integrated phylogenetic analysis. Strain relatedness was assessed by repetitive-sequence-based PCR (rep-PCR) and pulsed-field gel electrophoresis (PFGE), which showed the characteristic clonal groups for each species. Five isolates with ambiguous species identities as M. abscessus-M. massiliense by rpoB, hsp65, and secA sequencing clustered as a distinct group by rep-PCR and PFGE together with the M. massiliense type strain. Overall, the clinical manifestations of disease caused by each species were similar. In summary, a multilocus sequencing approach (not just rpoB partial sequencing) is required for division of M. abscessus and closely related species. Molecular typing complements sequence-based identification and provides information on prevalent clones with possible relevant clinical aspects.
Assuntos
Técnicas de Tipagem Bacteriana/métodos , Técnicas de Diagnóstico Molecular/métodos , Infecções por Mycobacterium/microbiologia , Mycobacterium/classificação , Mycobacterium/isolamento & purificação , Adenosina Trifosfatases/genética , Adolescente , Adulto , Idoso , Proteínas de Bactérias/genética , Chaperonina 60 , Chaperoninas/genética , Criança , Análise por Conglomerados , Estudos de Coortes , Impressões Digitais de DNA , DNA Bacteriano/química , DNA Bacteriano/genética , RNA Polimerases Dirigidas por DNA/genética , Eletroforese em Gel de Campo Pulsado , Feminino , Humanos , Masculino , Proteínas de Membrana Transportadoras/genética , Pessoa de Meia-Idade , Mycobacterium/genética , Filogenia , Reação em Cadeia da Polimerase/métodos , Canais de Translocação SEC , Proteínas SecA , Análise de Sequência de DNA , Adulto JovemRESUMO
Chronic granulomatous disease (CGD) is a rare inherited disease of the phagocyte NADPH oxidase system causing defective production of toxic oxygen metabolites, impaired bacterial and fungal killing, and recurrent life-threatening infections. We identified a novel gram-negative rod in excised lymph nodes from a patient with CGD. Gram-negative rods grew on charcoal-yeast extract, but conventional tests could not identify it. The best 50 matches of the 16S rRNA (using BLAST) were all members of the family Acetobacteraceae, with the closest match being Gluconobacter sacchari. Patient serum showed specific band recognition in whole lysate immunoblot. We used mouse models of CGD to determine whether this organism was a genuine CGD pathogen. Intraperitoneal injection of gp91(phox -/-) (X-linked) and p47 (phox -/-) (autosomal recessive) mice with this bacterium led to larger burdens of organism recovered from knockout compared with wild-type mice. Knockout mouse lymph nodes had histopathology that was similar to that seen in our patient. We recovered organisms with 16S rRNA sequence identical to the patient's original isolate from the infected mice. We identified a novel gram-negative rod from a patient with CGD. To confirm its pathogenicity, we demonstrated specific immune reaction by high titer antibody, showed that it was able to cause similar disease when introduced into CGD, but not wild-type mice, and we recovered the same organism from pathologic lesions in these mice. Therefore, we have fulfilled Koch's postulates for a new pathogen. This is the first reported case of invasive human disease caused by any of the Acetobacteraceae. Polyphasic taxonomic analysis shows this organism to be a new genus and species for which we propose the name Granulobacter bethesdensis.
Assuntos
Infecções Bacterianas/complicações , Bactérias Gram-Negativas/isolamento & purificação , Doença Granulomatosa Crônica/microbiologia , Linfadenite/microbiologia , Adulto , Sequência de Aminoácidos , Animais , Infecções Bacterianas/patologia , Infecções Bacterianas/terapia , Bactérias Gram-Negativas/classificação , Bactérias Gram-Negativas/genética , Doença Granulomatosa Crônica/patologia , Doença Granulomatosa Crônica/terapia , Humanos , Linfonodos/microbiologia , Linfonodos/patologia , Linfadenite/patologia , Linfadenite/terapia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , RNA Ribossômico 16S/genéticaRESUMO
We examined a cluster of 5 hemodialysis patients who contracted gram-negative bacteremia. A nurse who used an artificial fingernail to open a vial of heparin that was mixed to make a flush solution had a culture of an artificial fingernail specimen positive for Serratia marcescens. The typing of the S. marcescens strains isolated from the 5 patients and the nurse showed them to be identical. This finding provides strong support for policies prohibiting artificial nails for healthcare workers in all hemodialysis units.
Assuntos
Bacteriemia/transmissão , Infecção Hospitalar/transmissão , Transmissão de Doença Infecciosa do Profissional para o Paciente/métodos , Unhas/microbiologia , Diálise Renal/efeitos adversos , Infecções por Serratia/transmissão , Serratia marcescens/isolamento & purificação , Bacteriemia/microbiologia , Indústria da Beleza , Infecção Hospitalar/microbiologia , Impressões Digitais de DNA , Desinfecção das Mãos/métodos , Humanos , Controle de Infecções/métodos , Transmissão de Doença Infecciosa do Profissional para o Paciente/prevenção & controle , Infusões Intravenosas/efeitos adversos , Recursos Humanos de Enfermagem , Infecções por Serratia/microbiologia , Infecções por Serratia/prevenção & controleRESUMO
PURPOSE: A strain of Salmonella typhimurium (VNP20009), attenuated by chromosomal deletion of the purI and msbB genes, was found to target to tumor and inhibit tumor growth in mice. These findings led to the present phase I study of the intravenous infusion of VNP20009 to patients with metastatic cancer. PATIENTS AND METHODS: In cohorts consisting of three to six patients, 24 patients with metastatic melanoma and one patient with metastatic renal cell carcinoma received 30-minute intravenous bolus infusions containing 10(6) to 10(9) cfu/m(2) of VNP20009. Patients were evaluated for dose-related toxicities, selective replication within tumors, and antitumor effects. RESULTS: The maximum-tolerated dose was 3 x 10(8) cfu/m(2). Dose-limiting toxicity was observed in patients receiving 1 x 10(9) cfu/m(2), which included thrombocytopenia, anemia, persistent bacteremia, hyperbilirubinemia, diarrhea, vomiting, nausea, elevated alkaline phosphatase, and hypophosphatemia. VNP20009 induced a dose-related increase in the circulation of proinflammatory cytokines, such as interleukin (IL)-1beta, tumor necrosis factor alpha, IL-6, and IL-12. Focal tumor colonization was observed in two patients receiving 1 x 10(9) cfu/m(2) and in one patient receiving 3 x 10(8) cfu/m(2). None of the patients experienced objective tumor regression, including those patients with colonized tumors. CONCLUSION: The VNP20009 strain of Salmonella typhimurium can be safely administered to patients, and at the highest tolerated dose, some tumor colonization was observed. No antitumor effects were seen, and additional studies are required to reduce dose-related toxicity and improve tumor localization.
Assuntos
Antineoplásicos/uso terapêutico , Carcinoma de Células Renais/secundário , Carcinoma de Células Renais/terapia , Melanoma/secundário , Melanoma/terapia , Salmonella typhimurium , Adulto , Idoso , Anticorpos Antibacterianos/sangue , Antineoplásicos/efeitos adversos , Translocação Bacteriana , Carcinoma de Células Renais/microbiologia , Carcinoma de Células Renais/patologia , Contagem de Colônia Microbiana , Citocinas/sangue , Relação Dose-Resposta Imunológica , Feminino , Engenharia Genética , Humanos , Infusões Intravenosas , Masculino , Dose Máxima Tolerável , Melanoma/microbiologia , Melanoma/patologia , Pessoa de Meia-Idade , Salmonella typhimurium/genética , Salmonella typhimurium/imunologiaRESUMO
The first described, environmentally isolated, Bordetella petrii was shown to undergo massive genomic rearrangements in vitro. More recently, B. petrii was isolated from clinical samples associated with jaw, ear bone, cystic fibrosis and chronic pulmonary disease. However, the in vivo consequences of B. petrii genome plasticity and its pathogenicity remain obscure. B. petrii was identified from four sequential respiratory samples and a post-mortem spleen sample of a woman presenting with bronchiectasis and cavitary lung disease associated with nontuberculous mycobacterial infection. Strains were compared genetically, phenotypically and by antibody recognition from the patient and from inoculated mice. The successive B. petrii strains exhibited differences in growth, antibiotic susceptibility and recognition by the patient's antibodies. Antibodies from mice inoculated with these strains recapitulated the specificity and strain dependent response that was seen with the patient's serum. Finally, we characterize one strain that was poorly recognized by the patient's antibodies, due to a defect in the lipopolysaccharide O-antigen, and identify a mutation associated with this phenotype. We propose that B. petrii is remarkably adaptable in vivo, providing a possible connection between immune response and bacterial evasion and supporting infection persistence.
Assuntos
Adaptação Fisiológica , Bordetella/fisiologia , Imunidade Adaptativa , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Bordetella/genética , Bordetella/imunologia , Feminino , Humanos , Evasão da Resposta Imune , Imunização , Pneumopatias/sangue , Pneumopatias/imunologia , Pneumopatias/microbiologia , Camundongos , Pessoa de Meia-Idade , Mutação , Antígenos O/genética , Análise de SequênciaRESUMO
The Gram-negative bacteria Klebsiella pneumoniae is a major cause of nosocomial infections, primarily among immunocompromised patients. The emergence of strains resistant to carbapenems has left few treatment options, making infection containment critical. In 2011, the U.S. National Institutes of Health Clinical Center experienced an outbreak of carbapenem-resistant K. pneumoniae that affected 18 patients, 11 of whom died. Whole-genome sequencing was performed on K. pneumoniae isolates to gain insight into why the outbreak progressed despite early implementation of infection control procedures. Integrated genomic and epidemiological analysis traced the outbreak to three independent transmissions from a single patient who was discharged 3 weeks before the next case became clinically apparent. Additional genomic comparisons provided evidence for unexpected transmission routes, with subsequent mining of epidemiological data pointing to possible explanations for these transmissions. Our analysis demonstrates that integration of genomic and epidemiological data can yield actionable insights and facilitate the control of nosocomial transmission.
Assuntos
Carbapenêmicos/farmacologia , Infecção Hospitalar/epidemiologia , Surtos de Doenças/estatística & dados numéricos , Farmacorresistência Bacteriana/genética , Genoma Bacteriano/genética , Klebsiella pneumoniae/genética , Análise de Sequência de DNA/métodos , Adulto , Idoso , Colistina/farmacologia , Busca de Comunicante , Infecção Hospitalar/microbiologia , Infecção Hospitalar/transmissão , Farmacorresistência Bacteriana/efeitos dos fármacos , Feminino , Hospitais/estatística & dados numéricos , Humanos , Infecções por Klebsiella/epidemiologia , Infecções por Klebsiella/microbiologia , Infecções por Klebsiella/transmissão , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/isolamento & purificação , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Mutação/genética , Polimorfismo de Nucleotídeo Único/genética , Estados Unidos/epidemiologia , Adulto JovemRESUMO
BACKGROUND: Multidrug-resistant Acinetobacter baumannii (MDRAB) is difficult to treat and eradicate. Several reports describe isolation and environmental cleaning strategies that controlled hospital MDRAB outbreaks. Such interventions were insufficient to interrupt MDRAB transmission in 2 intensive care unit-based outbreaks in our hospital. We describe strategies that were associated with termination of MDRAB outbreaks at the National Institutes of Health Clinical Center. METHODS: In response to MDRAB outbreaks in 2007 and 2009, we implemented multiple interventions, including stakeholder meetings, enhanced isolation precautions, active microbial surveillance, cohorting, and extensive environmental cleaning. We conducted a case-control study to analyze risk factors for acquiring MDRAB. In each outbreak, infection control adherence monitors were placed in MDRAB cohort areas to observe and correct staff infection control behavior. RESULTS: Between May 2007 and December 2009, 63 patients acquired nosocomial MDRAB; 57 (90%) acquired 1 or more of 4 outbreak strains. Of 347 environmental cultures, only 2 grew outbreak strains of MDRAB from areas other than MDRAB patient rooms. Adherence monitors recorded 1,330 isolation room entries in 2007, of which 8% required interventions. In 2009, around-the-clock monitors recorded 4,892 staff observations, including 127 (2.6%) instances of nonadherence with precautions, requiring 68 interventions (1.4%). Physicians were responsible for more violations than other staff (58% of hand hygiene violations and 37% of violations relating to gown and glove use). Each outbreak terminated in temporal association with initiation of adherence monitoring. CONCLUSIONS: Although labor intensive, adherence monitoring may be useful as part of a multifaceted strategy to limit nosocomial transmission of MDRAB.
Assuntos
Infecções por Acinetobacter/epidemiologia , Infecções por Acinetobacter/prevenção & controle , Acinetobacter baumannii , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/prevenção & controle , Controle de Infecções/métodos , Infecções por Acinetobacter/tratamento farmacológico , Acinetobacter baumannii/efeitos dos fármacos , Acinetobacter baumannii/isolamento & purificação , Estudos de Casos e Controles , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/microbiologia , Surtos de Doenças/prevenção & controle , Farmacorresistência Bacteriana Múltipla , Fidelidade a Diretrizes/estatística & dados numéricos , Humanos , Unidades de Terapia Intensiva , Modelos Logísticos , Maryland/epidemiologia , National Institutes of Health (U.S.) , Enfermeiras e Enfermeiros , Roupa de Proteção/estatística & dados numéricos , Fatores de Risco , Vigilância de Evento Sentinela , Estados Unidos/epidemiologiaRESUMO
BACKGROUND: Nosocomial outbreaks of Legionnaires disease have been linked to contaminated water in hospitals. Immunocompromised patients are particularly vulnerable and, when infected, have a high mortality rate. We report the investigation of a cluster of cases of nosocomial pneumonia attributable to Legionella pneumophila serogroup 1 that occurred among patients on our stem cell transplantation unit. METHODS: We conducted a record review to identify common points of potential exposure, followed by environmental and water sampling for Legionella species from those sources. We used an air sampler to in an attempt to detect aerosolized Legionella and pulsed-field gel electrophoresis to compare clinical and environmental isolates. RESULTS: The most likely sources identified were the water supply in the patients' rooms and a decorative fountain in the radiation oncology suite. Samples from the patients' rooms did not grow Legionella species. Cultures of the fountain, which had been restarted 4 months earlier after being shut off for 5 months, yielded L. pneumophila serogroup 1. The isolates from both patients and the fountain were identical by pulsed-field gel electrophoresis. Both patients developed pneumonia within 10 days of completing radiation therapy, and each reported having observed the fountain at close range. Both patients' infections were identified early and treated promptly, and both recovered. CONCLUSIONS: This cluster was caused by contamination of a decorative fountain despite its being equipped with a filter and ozone generator. Fountains are a potential source of nosocomial Legionnaires disease despite standard maintenance and sanitizing measures. In our opinion, fountains present unacceptable risk in hospitals serving immunocompromised patients.
Assuntos
Infecção Hospitalar/epidemiologia , Infecção Hospitalar/transmissão , Doença dos Legionários/epidemiologia , Doença dos Legionários/transmissão , Microbiologia da Água , Adulto , Análise por Conglomerados , Infecção Hospitalar/microbiologia , Contaminação de Equipamentos , Unidades Hospitalares , Humanos , Hospedeiro Imunocomprometido , Legionella pneumophila/classificação , Legionella pneumophila/isolamento & purificação , Doença dos Legionários/microbiologia , MasculinoRESUMO
In this study, we investigated the clonal emergence of daptomycin-resistant Enterococcus faecium strains isolated from a patient with leukocyte adhesion deficiency syndrome. The resistance mechanism in these strains is independent of either equivalent point mutations previously described for Staphylococcus aureus or daptomycin inactivation mechanisms identified in soil bacteria.
Assuntos
Antibacterianos/farmacologia , Daptomicina/farmacologia , Farmacorresistência Bacteriana , Enterococcus faecium/efeitos dos fármacos , Infecções por Bactérias Gram-Positivas/microbiologia , Síndrome da Aderência Leucocítica Deficitária/microbiologia , Proteínas de Bactérias/genética , DNA Bacteriano/análise , DNA Bacteriano/isolamento & purificação , Farmacorresistência Bacteriana/genética , Eletroforese em Gel de Campo Pulsado , Enterococcus faecium/genética , Enterococcus faecium/isolamento & purificação , Humanos , Testes de Sensibilidade Microbiana , Mutação Puntual , Reação em Cadeia da Polimerase , Vancomicina/farmacologiaRESUMO
A Gram-negative, aerobic, coccobacillus to rod-shaped bacterium was isolated from three patients with chronic granulomatous disease. The organism was subjected to a polyphasic taxonomic study. A multilocus phylogenetic analysis based on the 16S rRNA gene, the internal transcribed spacer (ITS) region and the RecA protein demonstrated that the organism belongs to a new sublineage within the acetic acid bacteria in the family Acetobacteraceae. Phenotypic features are summarized as follows: the organism grew at an optimum temperature of 35-37 degrees C and optimum pH of 5.0-6.5. It produced a yellow pigment, oxidized lactate and acetate, the latter weakly, produced little acetic acid from ethanol and could use methanol as a sole carbon source. The two major fatty acids were a straight-chain unsaturated acid (C18:1omega7c) and C16:0. The DNA base composition was 59.1 mol% G+C. The very weak production of acetic acid from ethanol, the ability to use methanol, the yellow pigmentation and high optimum temperature for growth distinguished this organism from other acetic acid bacteria. The unique phylogenetic and phenotypic characteristics suggest that the bacterium should be classified within a separate genus, for which the name Granulibacter bethesdensis gen. nov., sp. nov. is proposed. The type strain is CGDNIH1T (=ATCC BAA-1260T=DSM 17861T).
Assuntos
Acetobacteraceae/classificação , Acetobacteraceae/isolamento & purificação , Doença Granulomatosa Crônica/microbiologia , Acetatos/metabolismo , Acetobacteraceae/citologia , Acetobacteraceae/fisiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico , Etanol/metabolismo , Ácidos Graxos/análise , Ácidos Graxos/química , Genes de RNAr , Humanos , Concentração de Íons de Hidrogênio , Ácido Láctico/metabolismo , Metanol/metabolismo , Dados de Sequência Molecular , Oxirredução , Filogenia , Pigmentos Biológicos/biossíntese , RNA Ribossômico 16S/genética , Recombinases Rec A/genética , Análise de Sequência de DNA , Temperatura , Estados UnidosRESUMO
Chronic granulomatous disease (CGD) is characterized by severe recurrent infections with Staphylococcus aureus, certain gram-negative rods, Nocardia species, and fungi. When infections with the same species recur, they may represent relapses or new infections. We collected organisms from infections that occurred between 1992 and 2000 in patients with CGD and determined the biochemical phenotypes, in vitro antibiotic susceptibility patterns, and pulsed-field gel electrophoresis (PFGE) patterns of the organisms causing the initial and recurrent infections. Recurrence of infection with Burkholderia cepacia or Serratia marcescens was caused by a new strain in 9 of 10 cases (P=.001). Recurrent S. aureus infections were caused by new strains in 7 of 8 cases (P=.006). In patients with CGD, recurrence of infection with the same bacterial species after appropriate antibiotic therapy usually represents new infection.