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1.
J Infect Chemother ; 29(8): 731-743, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37116613

RESUMO

The trends and prevalence of antimicrobial susceptibility of pathogens vary by country, region, and time. Long-term regular surveillance is required to investigate trends in the antimicrobial resistance of various isolated bacterial pathogens. We report the results of a nationwide surveillance on the antimicrobial susceptibility of bacterial respiratory pathogens in Japan conducted by the Japanese Society of Chemotherapy, the Japanese Association for Infectious Diseases, and the Japanese Society for Clinical Microbiology. The isolates were collected from clinical specimens obtained from adult patients who visited a collaborating medical facility between June 2019 and December 2020 and were diagnosed with respiratory tract infections by a physician. Antimicrobial susceptibility testing was performed in a centralized laboratory according to the methods recommended by the Clinical and Laboratory Standards Institute. Susceptibility testing was performed for 932 strains (201 Staphylococcus aureus, 158 Streptococcus pneumoniae, 6 S. pyogenes, 136 Haemophilus influenzae, 127 Moraxella catarrhalis, 141 Klebsiella pneumoniae, and 163 Pseudomonas aeruginosa) collected from 32 facilities in Japan. The proportions of methicillin-resistant S. aureus and penicillin-resistant S. pneumoniae were 35.3% and 0%, respectively. In H. influenzae, 16.2% and 16.9% were ß-lactamase-producing ampicillin resistant and ß-lactamase-negative ampicillin resistant, respectively. Extended-spectrum ß-lactamase-producing K. pneumoniae accounted for 5.0% of all K. pneumoniae infections. Carbapenemase-producing K. pneumoniae and multi-drug-resistant P. aeruginosa with metallo-ß-lactamase were not detected in this study. This surveillance will be a useful reference for treating respiratory infections in Japan and will provide evidence to enhance the appropriate use of antimicrobial agents.


Assuntos
Doenças Transmissíveis , Staphylococcus aureus Resistente à Meticilina , Infecções Respiratórias , Adulto , Humanos , Ampicilina , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Bactérias , beta-Lactamases , Doenças Transmissíveis/tratamento farmacológico , Farmacorresistência Bacteriana , Haemophilus influenzae , Testes de Sensibilidade Microbiana , Infecções Respiratórias/tratamento farmacológico , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/microbiologia , Japão
2.
Front Microbiol ; 12: 675463, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34054783

RESUMO

The rapid emergence of carbapenemase-producing gram-negative bacteria (CPGNB) is a global threat due to the high mortality of infection and limited treatment options. Although there have been many reports of CPGNB isolated from Southeast Asian countries, to date there has been no genetic analysis of CPGNB isolated from Cambodia. Sequence-based molecular epidemiological analysis enables a better understanding of the genotypic characteristics and epidemiological significance of antimicrobial-resistant (AMR) bacteria in each country, and allows countries to enact measures related to AMR issues. In this study, we performed on-site genomic epidemiological analysis of CPGNB isolated in Cambodia using a portable laboratory equipment called Bento Lab, which combines a PCR thermal cycler, microcentrifuge, gel electrophoresis apparatus, and LED transilluminator, along with the MinION nanopore sequencer. PCR targeting of major carbapenemase genes using Bento Lab revealed that two Escherichia coli isolates and one Acinetobacter baumannii isolate harbored carbapenemase genes: bla NDM, bla OXA-48, and bla OXA-23, respectively. The results of phenotypic diagnostic tests for CPGNB, such as the carbapenem inactivation method and double-disk diffusion test using a specific inhibitor of metallo-ß-lactamases, were consistent with their AMR genotypes. Whole-genome sequencing analysis using MinION revealed that bla NDM-5 gene was carried on a 93.9-kb plasmid with IncFIA/IncFIB/IncFII/IncQ1 replicons, and bla OXA-181 gene was carried on a 51.5-kb plasmid with the IncX3 replicon in E. coli isolates. bla OXA-23 was encoded in two locations on the chromosome of A. baumannii. Plasmids carrying bla NDM-5 or bla OXA-181 in E. coli were highly structurally identical to plasmids prevalent in Enterobacterales in China and other countries, suggesting that they disseminated from a common evolutionary origin. Our findings demonstrate the potential impact of portable laboratory equipment on AMR bacteria research in hospitals and research centers with limited research facilities, and provide the first glimpse into the genomic epidemiology of CPGNB in Cambodia.

3.
BMC Microbiol ; 18(1): 213, 2018 12 13.
Artigo em Inglês | MEDLINE | ID: mdl-30545294

RESUMO

BACKGROUND: VanB-type vancomycin (VAN) resistance gene clusters confer VAN resistances on Enterococcus spp. over a wide range of MIC levels (MIC = 4-1000 mg/L). However, the epidemiology and the molecular characteristics of the VAN susceptible VanB-type Enterococcus still remain unclear. RESULTS: We characterized 19 isolates of VanB-type Enterococcus faecium that might colonize in the gut and were not phenotypically resistant to VAN (MIC = 3 mg/L). They were obtained from two hospitals in Japan between 2009 and 2010. These isolates had the identical vanB gene cluster and showed same multilocus sequence typing (MLST) (ST78) and the highly related profiles in pulsed-field gel electrophoresis (PFGE). The vanB gene cluster was located on a plasmid, and was transferable to E. faecium and E. faecalis. Notably, from these VanB-type VREs, VAN resistant (MIC≥16 mg/L) mutants could appear at a frequency of 10- 6-10- 7/parent cell in vitro. Most of these revertants acquired mutations in the vanSB gene, while the remainder of the revertants might have other mutations outside of the vanB gene cluster. All of the revertants we tested showed increases in the VAN-dependent expression of the vanB gene cluster, suggesting that the mutations affected the transcriptional activity and increased the VAN resistance. Targeted mutagenesis revealed that three unique nucleotide substitutions in the vanB gene cluster of these strains attenuated VAN resistance. CONCLUSIONS: In summary, this study indicated that stealthy VanB-type E. faecium strains that have the potential ability to become resistance to VAN could exist in clinical settings.


Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Enterococcus faecium/efeitos dos fármacos , Enterococcus faecium/genética , Infecções por Bactérias Gram-Positivas/microbiologia , Proteínas de Bactérias/metabolismo , Eletroforese em Gel de Campo Pulsado , Enterococcus faecium/classificação , Enterococcus faecium/isolamento & purificação , Humanos , Japão , Família Multigênica , Tipagem de Sequências Multilocus , Mutação , Vancomicina/farmacologia , Resistência a Vancomicina
4.
Can J Infect Dis Med Microbiol ; 2017: 4896791, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28316631

RESUMO

Background. Early detection of Gram-positive bacteremia and timely appropriate antimicrobial therapy are required for decreasing patient mortality. The purpose of our study was to evaluate the performance of the Verigene Gram-positive blood culture assay (BC-GP) in two special healthcare settings and determine the potential impact of rapid blood culture testing for Gram-positive bacteremia within the Japanese healthcare delivery system. Furthermore, the study included simulated blood cultures, which included a library of well-characterized methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) isolates reflecting different geographical regions in Japan. Methods. A total 347 BC-GP assays were performed on clinical and simulated blood cultures. BC-GP results were compared to results obtained by reference methods for genus/species identification and detection of resistance genes using molecular and MALDI-TOF MS methodologies. Results. For identification and detection of resistance genes at two clinical sites and simulated blood cultures, overall concordance of BC-GP with reference methods was 327/347 (94%). The time for identification and antimicrobial resistance detection by BC-GP was significantly shorter compared to routine testing especially at the cardiology hospital, which does not offer clinical microbiology services on weekends and holidays. Conclusion. BC-GP generated accurate identification and detection of resistance markers compared with routine laboratory methods for Gram-positive organisms in specialized clinical settings providing more rapid results than current routine testing.

5.
J Infect Chemother ; 21(12): 849-56, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26433422

RESUMO

The Verigene Gram-positive blood culture test (BC-GP) and the Verigene Gram-negative blood culture test (BC-GN) identify representative Gram-positive bacteria, Gram-negative bacteria and their antimicrobial resistance by detecting resistance genes within 3 h. Significant benefits are anticipated due to their rapidity and accuracy, however, their clinical utility is unproven in clinical studies. We performed a clinical trial between July 2014 and December 2014 for hospitalized bacteremia patients. During the intervention period (N = 88), Verigene BC-GP and BC-GN was used along with conventional microbiological diagnostic methods, while comparing the clinical data and outcomes with those during the control period (N = 147) (UMIN registration ID: UMIN000014399). The median duration between the initiation of blood culture incubation and the reporting time of the Verigene system results was 21.7 h (IQR 18.2-26.8) and the results were found in 88% of the cases by the next day after blood cultures were obtained without discordance. The hospital-onset infection rate was higher in the control period (24% vs. 44%, p = 0.002), however, no differences were seen in co-morbidities and severity between the control and intervention periods. During the intervention period, the time of appropriate antimicrobial agents' initiation was significantly earlier than that in the control period (p = 0.001) and most cases (90%; 79/88) were treated with antimicrobial agents with in-vitro susceptibility for causative bacteria the day after the blood culture was obtained. The costs for antimicrobial agents were lower in the intervention period (3618 yen vs. 8505 yen, p = 0.001). The 30-day mortality was lower in the intervention period (3% vs. 13%, p = 0.019).


Assuntos
Bacteriemia/diagnóstico , Bacteriemia/microbiologia , Técnicas de Diagnóstico Molecular/instrumentação , Idoso , Antibacterianos/uso terapêutico , Bacteriemia/tratamento farmacológico , Doenças Transmissíveis/diagnóstico , Doenças Transmissíveis/tratamento farmacológico , Doenças Transmissíveis/microbiologia , Infecção Hospitalar/diagnóstico , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/microbiologia , Feminino , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/genética , Bactérias Gram-Positivas/efeitos dos fármacos , Bactérias Gram-Positivas/genética , Humanos , Masculino , Análise em Microsséries/métodos , Estudos Prospectivos
6.
Diagn Microbiol Infect Dis ; 83(4): 344-8, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26361710

RESUMO

The Verigene Gram-Negative Blood Culture Nucleic Acid Test (BC-GN) is a microarray-based assay that enables rapid detection of 9 common Gram-negative bacteria and 6 resistance determinants directly from positive blood cultures. We compared the performance of BC-GN with currently used automated systems, testing 141 clinical blood cultures and 205 spiked blood cultures. For identification of BC-GN target organisms in clinical and spiked blood cultures, the BC-GN assay showed 98.5% (130/132) and 98.9% (182/184) concordance, respectively. Of 140 resistance genes positively detected in clinical and spiked blood cultures with the BC-GN test, 139 (99.3%) were confirmed by PCR, and the detection results were consistent with the resistance phenotypes observed. The BC-GN assay, thus, can potentially improve care for sepsis patients by enabling timely detection and targeted antimicrobial therapy.


Assuntos
Bacteriemia/diagnóstico , Bacteriemia/microbiologia , DNA Bacteriano/análise , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/isolamento & purificação , Testes de Sensibilidade Microbiana/métodos , Técnicas de Diagnóstico Molecular/métodos , Automação Laboratorial/métodos , DNA Bacteriano/genética , Bactérias Gram-Negativas/classificação , Humanos , Fatores de Tempo
7.
Microb Drug Resist ; 21(4): 441-7, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25789579

RESUMO

Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) is now endemic in the United States. In Japan, CA-MRSA infections and CA-MRSA surveillance have been scarcely reported. In this study, we conducted a nationwide survey of CA-MRSA in Japan. We collected MRSA strains isolated from outpatients with skin and soft-tissue infection (SSTI) at 107 medical facilities from 24 prefectures in 2010 and 2012. Among 10,385 clinical samples from SSTI patients, 3,581 S. aureus isolates (35%) were obtained and 673 of the S. aureus strains (19%) were identified as MRSA. Among 625 MRSA strains tested in this study, 266 strains (43%) and 114 strains (18%) were classified as SCCmec types IV and V, respectively. Detection of virulence genes was as follows: Panton-Valentine leukocidin (PVL) gene (57 strains, 9%), exfoliative toxin (ET) gene (179 strains, 29%), toxic shock syndrome toxin-1 (TSST-1) gene (195 strains, 31%), or none. PVL-positive strains were classified into eight sequence types (STs) (i.e., ST1, ST5, ST8, ST22, ST30, ST452, ST59, and ST154) and six clonal complexes (i.e., CC1, CC5, CC8, CC22, CC30, and CC59). Only 10 PVL-positive strains (2%) were pulsed-field type USA300 clone. There were a wide variety of CA-MRSA clones in Japan, which were different from the situation in the United States.


Assuntos
Farmacorresistência Bacteriana/genética , Staphylococcus aureus Resistente à Meticilina/genética , Dermatopatias Infecciosas/microbiologia , Infecções Estafilocócicas/microbiologia , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Japão/epidemiologia , Masculino , Staphylococcus aureus Resistente à Meticilina/patogenicidade , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Pacientes Ambulatoriais , Reação em Cadeia da Polimerase , Vigilância em Saúde Pública , Pele/microbiologia , Dermatopatias Infecciosas/epidemiologia , Infecções dos Tecidos Moles/microbiologia , Infecções Estafilocócicas/epidemiologia , Supuração/microbiologia , Virulência , Adulto Jovem
8.
Microb Drug Resist ; 21(2): 224-33, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25419619

RESUMO

The rate of fluoroquinolone (FQ) resistance among the cephalosporin-resistant Klebsiella pneumoniae is considerably high, however, their genetic profiles have not been well investigated. We selected 61 ciprofloxacin-nonsusceptible isolates from 102 K. pneumoniae isolates judged to be "resistant" to some cephalosporins during 2009 and 2012 throughout Japan. Pulsed-field gel electrophoresis excluded clonal isolates, and 29 isolates were subjected to multilocus sequence typing (MLST), detection of the amino acid substitutions in the quinolone resistance determining regions (QRDRs) of GyrA and ParC, ß-lactamase typing, and identification of plasmid-mediated quinolone resistance (PMQR) genes. PCR-based replicon typing was performed, after PMQR gene transfer. Four major sequence types (STs) or clonal complexes (CCs), that is, ST37, CC17 (consisting of ST17 and ST20), ST11, and CC528 (consisting of ST528 and ST1130), were found, and they accounted for 48.2% of the isolates tested. Amino acid substitutions in the QRDRs and the presence of PMQR genes were identified in 20 (68.9%) and 18 (62.0%) isolates, respectively. The replicon type of three PMQR-carrying plasmids was IncN, but others were nontypable. Fifteen (83.3%) of the 18 PMQR-harboring isolates coharbored blaCTX-M and/or blaDHA-1. Ciprofloxacin-nonsusceptible K. pneumoniae clinical isolates demonstrating cephalosporin resistance often belong to the global epidemic lineages and possess PMQR and/or QRDR substitutions.


Assuntos
Cefalosporinas/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Fluoroquinolonas/farmacologia , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/genética , Substituição de Aminoácidos/genética , Antibacterianos/farmacologia , Ciprofloxacina/farmacologia , Genes Bacterianos/genética , Japão , Quinolonas/farmacologia , beta-Lactamases/genética
9.
BMC Infect Dis ; 14: 54, 2014 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-24484470

RESUMO

BACKGROUND: The rapid identification of acid-fast bacilli recovered from patient specimens as Mycobacterium tuberculosis complex (MTC) is critically important for accurate diagnosis and treatment. A thin-layer immunochromatographic (TLC) assay using anti-MPB64 or anti-MPT64 monoclonal antibodies was developed to discriminate between MTC and non-tuberculosis mycobacteria (NTM). Capilia TB-Neo, which is the improved version of Capilia TB, is recently developed and needs to be evaluated. METHODS: Capilia TB-Neo was evaluated by using reference strains including 96 Mycobacterium species (4 MTC and 92 NTM) and 3 other bacterial genera, and clinical isolates (500 MTC and 90 NTM isolates). M. tuberculosis isolates tested negative by Capilia TB-Neo were sequenced for mpt64 gene. RESULTS: Capilia TB-Neo showed 100% agreement to a subset of reference strains. Non-specific reaction to M. marinum was not observed. The sensitivity and specificity of Capilia TB-Neo to the clinical isolates were 99.4% (99.6% for M. tuberculosis, excluding M. bovis BCG) for clinical MTC isolates and 100% for NTM isolates tested, respectively. Two M. tuberculosis isolates tested negative by Capilia TB-Neo: one harbored a 63-bp deletion in the mpt64 gene and the other possessed a 3,659-bp deletion from Rv1977 to Rv1981c, a region including the entire mpt64 gene. CONCLUSIONS: Capilia TB-Neo is a simple, rapid and highly sensitive test for identifying MTC, and showed better specificity than Capilia TB. However, Capilia TB-Neo still showed false-negative results with mpt64 mutations. The limitation should be recognized for clinical use.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Cromatografia de Afinidade/métodos , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose/microbiologia , Proteínas de Bactérias/genética , Humanos , Mycobacterium tuberculosis/classificação , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/crescimento & desenvolvimento , Sensibilidade e Especificidade , Tuberculose/diagnóstico
10.
Jpn J Infect Dis ; 66(6): 546-8, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24270149

RESUMO

Streptococcus agalactiae (group B Streptococcus, GBS) is the leading cause of neonatal sepsis and meningitis and an important pathogen in elderly patients and those with underlying diseases. The diagnosis of GBS infections is primarily based on culture of GBS. Some clinical laboratories perform the Christie-Atkins-Munch-Peterson (CAMP) test for discrimination of GBS from other streptococci. Here, we developed a rapid GBS identification method, i.e., the loop-mediated isothermal amplification (LAMP) method for detecting the cfb gene encoding the CAMP factor. This method detected at least 4 copies of the cfb gene in GBS under isothermal conditions with in a short time (65 °C, within 90 min). No inappropriate amplification of nucleotide by this method was observed when the chromosomal DNA of 17 streptococci and enterococci species, other than GBS, were used as templates. In this investigation, we successfully developed a LAMP method for rapid and highly sensitive detection of GBS, which provides a beneficial alternative to the conventional CAMP test.


Assuntos
Técnicas de Amplificação de Ácido Nucleico/métodos , Infecções Estreptocócicas/microbiologia , Streptococcus agalactiae/isolamento & purificação , Humanos , Tipagem Molecular/métodos , Infecções Estreptocócicas/diagnóstico , Streptococcus agalactiae/genética
11.
J Clin Microbiol ; 51(6): 1762-8, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23536409

RESUMO

Metallo-ß-lactamases (MBLs) are transmissible carbapenemases of increasing prevalence in Gram-negative bacteria among health care facilities worldwide. Control of the further spread of these carbapenem-resistant bacteria relies on clinical microbiological laboratories correctly identifying and classifying the MBLs. In this study, we evaluated a simple and rapid method for detecting IMP, the most prevalent MBL in Japan. We used an immunochromatography (IC) assay for 181 carbapenem-nonsusceptible (CNS) (nonsusceptible to imipenem or meropenem) strains comprising 74 IMP-producing and 33 non-IMP-producing strains of non-glucose-fermenting Gram-negative rods (NFGNR), as well as 64 IMP-producing and 10 non-IMP-producing Enterobacteriaceae strains. The IC assay results were compared to those from the double-disk synergy test (DDST), the MBL Etest, and the modified Hodge test (MHT) (only for Enterobacteriaceae). The IMP type was confirmed by specific PCR and direct sequencing. The IC assay detected all of the IMP-type MBLs, including IMP-1, -2, -6, -7, -10, -11, -19, -20, and -22 and IMP-40, -41, and -42 (new types), with 100% specificity and sensitivity against all strains tested. Although the sensitivity and specificity values for the DDST and MHT were equivalent to those for the IC assay, the MBL Etest was positive for only 87% of NFGNR and 31% of Enterobacteriaceae due to the low MIC of imipenem, causing an indeterminate evaluation. These results indicated that the IC assay might be a useful alternative to PCR for IMP MBL detection screening.


Assuntos
Antibacterianos/farmacologia , Carbapenêmicos/farmacologia , Cromatografia de Afinidade/métodos , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/enzimologia , Resistência beta-Lactâmica , beta-Lactamases/análise , Técnicas Bacteriológicas/métodos , Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Japão , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade , Análise de Sequência de DNA , beta-Lactamases/genética
12.
Case Rep Infect Dis ; 2012: 358789, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22924139

RESUMO

Central nervous system infections caused by Bacillus cereus have rarely been reported in infants. In this paper, the case of a 2-month-old low-birth-weight female who developed meningitis 45 days after resolution of a bloodstream infection (BSI) is described. The pulsed-field gel electrophoresis results revealed that the patterns of both B. cereus isolates responsible for the acute meningitis and for the prior bacteraemic episode were closely related. Although the source of the infection from within the patient was not clear, it is suggested that the B. cereus BSI developed in the neonate was complicated by acute meningitis.

13.
FEMS Immunol Med Microbiol ; 63(2): 202-9, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22077223

RESUMO

We characterized the profiles of virulence genes and antimicrobial susceptibility of Bacillus cereus isolates from blood cultures as well as the risk factors for blood stream infections (BSIs). The diversity of virulence gene patterns was found to be wide among 15 B. cereus isolates from BSIs and also among 11 isolates from contaminated blood cultures. The MicroScan broth microdilution method yielded results corresponding with those of the agar dilution (reference) method for levofloxacin, linezolid, and vancomycin, while the Etest results were consistent with the reference results for clindamycin, gentamicin, imipenem, levofloxacin, and linezolid. Compared with the reference values, however, some isolates showed marked differences of the minimum inhibitory concentrations (MICs) for ampicillin and clindamycin when determined using the MicroScan method, or the MICs for ampicillin, meropenem, and vancomycin when determined using the Etest method. Significantly more patients were treated with antimicrobials for more than 3 days during the 3-month period before isolation in the BSI group. Prior antimicrobial therapy may be a risk factor for BSIs due to B. cereus.


Assuntos
Antibacterianos/farmacologia , Infecções por Bacillaceae/microbiologia , Bacillus cereus/isolamento & purificação , Bacteriemia/microbiologia , Sangue/microbiologia , Fatores de Virulência/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/uso terapêutico , Infecções por Bacillaceae/epidemiologia , Bacillus cereus/classificação , Bacillus cereus/efeitos dos fármacos , Bacillus cereus/genética , Bacteriemia/epidemiologia , Criança , Pré-Escolar , Análise por Conglomerados , Eletroforese em Gel de Campo Pulsado , Feminino , Variação Genética , Humanos , Lactente , Recém-Nascido , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Tipagem Molecular , Fatores de Risco , Adulto Jovem
14.
Infect Genet Evol ; 11(1): 52-6, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20969979

RESUMO

The genus Acinetobacter is an important causative pathogen of nosocomial infections in the healthcare setting. The objectives of this study were to determine the species of causative pathogens and the sources of Acinetobacter blood stream infections that occurred in 2 immunocompetent pregnant women admitted to an obstetrics ward within a 2-month period. Phenotypic identification of the two isolates from blood stream infections was inconsistent among the ID test, the MicroScan WalkAway and the Vitek2 systems. In addition to the growth profile and detailed biochemical analysis, genotypic identification and phylogenetic tree analysis based on the almost complete 16S rRNA sequence and the partial rpoB gene sequence confirmed the identification of these isolates as A. ursingii. Environmental investigation of the obstetrics ward revealed A. ursingii and different strains of Acinetobacter junii in specimens obtained from the ward shower bath, although the source and route of transmission for the A. ursingii infections were not clarified. Our findings show that A. ursingii can inhabit the hospital environment.


Assuntos
Acinetobacter/patogenicidade , Obstetrícia , Complicações Infecciosas na Gravidez/microbiologia , Sepse/microbiologia , Eletroforese em Gel de Campo Pulsado , Feminino , Humanos , Testes de Sensibilidade Microbiana , Filogenia , Gravidez , Sepse/complicações
15.
Microbiol Res ; 165(1): 11-20, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-18417334

RESUMO

Haemophilus influenzae is a common pathogen of respiratory infections. We examined whether beta-lactamase-negative ampicillin-resistant (BLNAR) strains that are known to have ampicillin resistance due to a substitution of amino acid of penicillin binding protein (PBP)-3, differ from beta-lactamase-negative ampicillin-susceptible strains with regard to invasion of bronchial epithelium. After 3h incubation of each of 34 beta-lactamase-negative ampicillin-susceptible and 57 BLNAR strains in the presence of BEAS-2B cells, a human bronchial epithelium cell line, extracellular bacteria were killed using gentamicin and intracellular bacteria numbered. All nine strains in which the efficiency of invasion was 1% or higher were BLNAR strains. The rate of invasion was significantly greater in strains with PBP-3 amino acid substitution (Met377 to Ile, Ser385 to Thr, Leu389 to Phe, and Asn526 to Lys) (n=34) than in those with no amino acid substitution. Electron microscopy showed that high invasive BLNAR strains were observed in cytoplasm of BEAS-2B cell layer. The injured cells were 9.44+/-1.76% among attaching cells examined by trypan blue staining after 6h. These data may suggest that the amino acid substitution of the PBP in BLNAR strains may at least partly play roles in macropinocytosis, leading to the invasion and injury to epithelial cells.


Assuntos
Substituição de Aminoácidos , Brônquios/microbiologia , Células Epiteliais/microbiologia , Infecções por Haemophilus/microbiologia , Haemophilus influenzae/metabolismo , Proteínas de Ligação às Penicilinas/genética , Ampicilina/farmacologia , Resistência a Ampicilina , Brônquios/citologia , Linhagem Celular , Haemophilus influenzae/efeitos dos fármacos , Haemophilus influenzae/genética , Haemophilus influenzae/fisiologia , Humanos , Mutação de Sentido Incorreto , Proteínas de Ligação às Penicilinas/metabolismo
16.
Rinsho Byori ; 57(5): 463-6, 2009 May.
Artigo em Japonês | MEDLINE | ID: mdl-19522253

RESUMO

With the revision of the Medical Service Law in 2006 by the Japanese Ministry of Health, Labour and Welfare (MHLW), all healthcare institutions are now required to implement a healthcare risk management program including infection control program. At a national level, an infection control surveillance program (JANIS) was implemented in July 2007. Regular weekly, monthly, and yearly infection control surveillance reports from independent microbiology laboratories can make significant contributions to infection control programs in small to mid-sized hospitals; furthermore, such programs are consistent with the framework of the MHLW's objective of strengthening risk management in healthcare institutions. Against the backdrop of current efforts to improve risk management, independent laboratories can make a significant contribution. Independent laboratories must play a role beyond merely receiving and processing specimens for microbiological examination. In addition to generating results for patients, hospital epidemiological data that contribute to local infection control programs must be a value-added component of the service. A major obstacle for independent laboratories to make a significant contribution to risk management is the current reimbursement system, which makes it economically impossible for independent laboratories to support infection control programs in healthcare institutions.


Assuntos
Infecção Hospitalar/prevenção & controle , Hospitais , Controle de Infecções , Laboratórios , Microbiologia , Serviços Terceirizados , Papel Profissional , Gestão da Segurança , Número de Leitos em Hospital , Humanos , Japão
17.
Antimicrob Agents Chemother ; 52(12): 4258-67, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18809936

RESUMO

The recent emergence of group B streptococcal isolates exhibiting increased penicillin MICs at the Funabashi Municipal Medical Center and other hospitals in Japan prompted a comparative analysis of the penicillin-binding proteins (PBPs) from those strains with the PBPs from penicillin-susceptible strains comprising four neonatal invasive strains isolated from 1976 to 1988 and two recent isolates. The PBP sequences of the penicillin-susceptible strains were highly conserved, irrespective of their isolation date. Of six strains with reduced susceptibility to penicillin (penicillin MICs, 0.25 to 0.5 mug/ml), strains R1, R2, R5, and R6 shared a unique set of five amino acid substitutions, including V405A adjacent to the (402)SSN(404) motif in PBP 2X and one in PBP 2B. The remaining two strains, R3 and R4, shared several substitutions, including Q557E adjacent to the (552)KSG(554) motif in PBP 2X, in addition to the substitutions in PBP 2B, which are commonly found among penicillin-insusceptible strains. Strains R7 and R8, which had a penicillin MIC of 1 mug/ml, shared a unique set of eight amino acid substitutions (two in PBP 2X; two in PBP 2B, including G613R adjacent to the (614)KTG(616) motif; three in PBP 1A; and one in PBP 2A), and the Q557E substitution in PBP 2X was common to R3 and R4. The binding of Bocillin FL was reduced or not detected in some PBPs, including PBP 2X of penicillin-insusceptible strains, but no significant reduction in the level of pbp2x transcription was found in such strains. The results of phylogenetic comparative analyses imply the absence of epidemic penicillin-insusceptible strains, and several genetic lineages of penicillin-insusceptible strains have been independently emerging through the accumulation of mutations in their pbp genes, especially in pbp2x.


Assuntos
Antibacterianos/farmacologia , Heterogeneidade Genética , Resistência às Penicilinas/genética , Proteínas de Ligação às Penicilinas/genética , Penicilinas/farmacologia , Streptococcus agalactiae/efeitos dos fármacos , Aminoaciltransferases/genética , Proteínas de Bactérias/genética , Feminino , Humanos , Recém-Nascido , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Proteínas de Ligação às Penicilinas/metabolismo , Filogenia , Análise de Sequência de DNA , Streptococcus agalactiae/genética
19.
Int J Antimicrob Agents ; 30(3): 255-8, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17616451

RESUMO

Antimicrobial therapy of Mycobacterium abscessus infection is difficult because there are relatively few effective treatment regimens and single-agent therapy frequently fails clinically. In light of the lack of data on the susceptibility profile of M. abscessus strains recovered from infections in Japan, the in vitro activity of imipenem in combination with clarithromycin, levofloxacin, moxifloxacin, minocycline, amikacin or tobramycin was investigated by the checkerboard method and compared with the combination amikacin and cefoxitin. The combination of imipenem with moxifloxacin, levofloxacin or clarithromycin had a higher synergistic and/or additive effect than amikacin and cefoxitin. A decrease in the MIC(90) value (minimum inhibitory concentration for 90% of the organisms) was observed in the presence of imipenem for clarithromycin, minocycline, levofloxacin and moxifloxacin. The data suggest that a combination regimen including imipenem may be a good choice in empirical treatment of M. abscessus infection.


Assuntos
Antibacterianos/farmacologia , Imipenem/farmacologia , Mycobacterium/efeitos dos fármacos , Sinergismo Farmacológico , Humanos , Testes de Sensibilidade Microbiana , Controle de Qualidade , Escarro/microbiologia , Infecção dos Ferimentos/microbiologia
20.
Jpn J Antibiot ; 55(5): 524-36, 2002 Oct.
Artigo em Japonês | MEDLINE | ID: mdl-12532636

RESUMO

We determined the minimum inhibitory concentration (MIC) of various antimicrobial agents against 108 strains of Streptococcus pneumoniae and 144 strains of Haemophilus influenzae isolated from respiratory organs in the First Department of Internal Medicine, Shinshu University, and affiliated hospitals between January 2000 and February 2001. The following results were obtained. 1. Fifty-one (47.2%), 56 (51.9%), and 1 (0.9%) of 108 strains of S. pneumoniae were classified as penicillin-susceptible S. pneumoniae (PSSP), penicillin-intermediate S. pneumoniae (PISP), and penicillin-resistant S. pneumoniae (PRSP), respectively. 2. Three carbapenems had potent antimicrobial activity against PISP and PRSP. Furthermore, none of the strains were highly resistant (MIC > 2 micrograms/ml) to benzylpenicillin, ampicillin (ABPC), sulbactam/ampicillin (SBT/ABPC), cefotaxime (CTX), or cefepime (CFPM). 3. Eleven (7.6%) and 6 (4.2%) of 144 strains of H. influenzae were classified as beta-lactamase-producing ABPC-resistant strains and beta-lactamase negative ABPC-resistant H. influenzae (BLNAR), respectively. Levofloxacin, sulfamethoxazole/trimethoprim, and meropenem had potent antimicrobial activity against these resistant strains. 4. BLNAR strains were more highly resistant to CTX, CFPM, SBT/ABPC, and cefaclor than beta-lactamaseproducing strains. 5. In our surveillance study regarding clinical isolates of S. pneumoniae and H. influenzae from respiratory organs in Nagano prefecture, there were regional differences in the isolation rate and antimicrobial susceptibility. The isolation rates of resistant strains were lower than those reported in a nationwide survey.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Haemophilus influenzae/efeitos dos fármacos , Streptococcus pneumoniae/efeitos dos fármacos , Resistência a Ampicilina , Resistência às Cefalosporinas , Haemophilus influenzae/isolamento & purificação , Humanos , Lactamas , Testes de Sensibilidade Microbiana , Resistência às Penicilinas , Infecções Respiratórias/microbiologia , Streptococcus pneumoniae/isolamento & purificação , Resistência beta-Lactâmica
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