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1.
High Blood Press Cardiovasc Prev ; 26(3): 217-225, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31236901

RESUMO

INTRODUCTION: A possible role of the oral microbiome, specifically oral nitrate reducing flora, in blood pressure (BP) homeostasis, if proven etiologic in nature, could lead to novel mechanism-based therapy to improve hypertension prevention and control. AIM: This cross-sectional study characterized and compared the oral microbiome between four study groups based on BP status among 446 postmenopausal women aged 53-82 years. METHODS: Three study groups were not taking hypertension medication and were separated based on BP, as follows: normal BP (systolic < 120 and diastolic < 80; N = 179), elevated BP/Stage I hypertension (systolic 120-139 or diastolic 80-90; N = 106), Stage II hypertension (systolic > 140 or diastolic > 90; N = 42). The forth group consisted of anyone taking hypertension medications, regardless of BP (N = 119). Subgingival microbiome composition was determined using 16S rRNA sequencing with the Illumina MiSeq platform. Kruskal-Wallis tests were used to compare species-level relative abundance of bacterial operational taxonomic units across the four groups. RESULTS: Sixty-five bacterial species demonstrated significant differences in relative abundance in women with elevated BP or using hypertension medication as compared to those with normal BP. After correction for multiple testing, two species, Prevotella oral (species 317) and Streptococcus oralis, remained significant and were lower in abundance among women taking antihypertension medications compared to those with normal BP (corrected P < 0.05). CONCLUSIONS: These data provide novel description of oral subgingival bacteria grouped according to BP status. Additional larger studies including functional analysis and prospective designs will help further assess the potential role of the oral microbiome in BP regulation and hypertension.


Assuntos
Bactérias/isolamento & purificação , Pressão Sanguínea , Hipertensão/microbiologia , Hipertensão/fisiopatologia , Microbiota , Boca/microbiologia , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Anti-Hipertensivos/uso terapêutico , Bactérias/classificação , Bactérias/genética , Pressão Sanguínea/efeitos dos fármacos , Estudos Transversais , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Hipertensão/diagnóstico , Hipertensão/tratamento farmacológico , Pessoa de Meia-Idade , Pós-Menopausa , Ribotipagem/métodos , Fatores de Risco , Fatores Sexuais
2.
Microb Genom ; 5(7)2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31107202

RESUMO

Clostridioides difficile is a ubiquitous, diarrhoeagenic pathogen often associated with healthcare-acquired infections that can cause a range of symptoms from mild, self-limiting disease to toxic megacolon and death. Since the early 2000s, a large proportion of C. difficile cases have been attributed to the ribotype 027 (RT027) lineage, which is associated with sequence type 1 (ST1) in the C. difficile multilocus sequence typing scheme. The spread of ST1 has been attributed, in part, to resistance to fluoroquinolones used to treat unrelated infections, which creates conditions ideal for C. difficile colonization and proliferation. In this study, we analysed 27 isolates from a healthcare network in northern Arizona, USA, and 1352 publicly available ST1 genomes to place locally sampled isolates into a global context. Whole genome, single nucleotide polymorphism analysis demonstrated that at least six separate introductions of ST1 were observed in healthcare facilities in northern Arizona over an 18-month sampling period. A reconstruction of transmission networks identified potential nosocomial transmission of isolates, which were only identified via whole genome sequence analysis. Antibiotic resistance heterogeneity was observed among ST1 genomes, including variability in resistance profiles among locally sampled ST1 isolates. To investigate why ST1 genomes are so common globally and in northern Arizona, we compared all high-quality C. difficile genomes and identified that ST1 genomes have gained and lost a number of genomic regions compared to all other C. difficile genomes; analyses of other toxigenic C. difficile sequence types demonstrate that this loss may be anomalous and could be related to niche specialization. These results suggest that a combination of antimicrobial resistance and gain and loss of specific genes may explain the prominent association of this sequence type with C. difficile infection cases worldwide. The degree of genetic variability in ST1 suggests that classifying all ST1 genomes into a quinolone-resistant hypervirulent clone category may not be appropriate. Whole genome sequencing of clinical C. difficile isolates provides a high-resolution surveillance strategy for monitoring persistence and transmission of C. difficile and for assessing the performance of infection prevention and control strategies.


Assuntos
Infecções por Clostridium/microbiologia , Infecções por Clostridium/transmissão , Clostridium difficile/isolamento & purificação , Infecção Hospitalar/microbiologia , Infecção Hospitalar/transmissão , Arizona , Infecções por Clostridium/prevenção & controle , Clostridium difficile/classificação , Clostridium difficile/genética , Infecção Hospitalar/prevenção & controle , DNA Bacteriano/genética , Genoma Bacteriano , Genômica , Humanos , Filogenia , Ribotipagem/métodos , Sequenciamento Completo do Genoma
3.
Ann Vasc Surg ; 60: 475.e11-475.e17, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31075452

RESUMO

BACKGROUND: Streptococcus pneumoniae is considered a rare cause of mycotic aneurysms. The microbiological diagnosis of mycotic aneurysms can be difficult, and many patients have negative blood culture results. METHODS: We describe a series of four consecutive cases of mycotic aneurysms caused by S. pneumoniae with no respiratory features or extravascular septic foci. In two patients with negative blood culture results, 16S PCR was used for the diagnosis of S. pneumoniae infection. RESULTS: Four men with mycotic aneurysms affecting the aorta, axillary, and popliteal arteries caused by S. pneumoniae presented to our center between 2015 and 2016. All were treated with at least one month of intravenous antibiotics, followed by at least 4 weeks of oral antibiotics. Two were additionally managed using endovascular surgical techniques, and one underwent an open surgical repair. The fourth patient presented with bilateral popliteal aneurysms, one of which ruptured and was managed using surgical ligation and bypass, whereas the other side subsequently ruptured and was repaired endovascularly. Three of the four patients are currently off antibiotics and considered cured, while one died of an unrelated cause. CONCLUSIONS: S. pneumoniae should be considered a potential causative agent of mycotic aneurysms. Diagnosis can be confirmed using 16S PCR, especially in patients where peripheral blood cultures are uninformative.


Assuntos
Aneurisma Infectado/microbiologia , Aneurisma Roto/microbiologia , Aneurisma Aórtico/microbiologia , DNA Bacteriano/genética , Aneurisma Ilíaco/microbiologia , Infecções Pneumocócicas/microbiologia , Reação em Cadeia da Polimerase , Ribotipagem/métodos , Streptococcus pneumoniae/genética , Idoso , Aneurisma Infectado/diagnóstico por imagem , Aneurisma Infectado/cirurgia , Aneurisma Roto/diagnóstico por imagem , Aneurisma Roto/cirurgia , Antibacterianos/uso terapêutico , Aneurisma Aórtico/diagnóstico por imagem , Aneurisma Aórtico/terapia , Humanos , Aneurisma Ilíaco/diagnóstico por imagem , Aneurisma Ilíaco/terapia , Masculino , Pessoa de Meia-Idade , Infecções Pneumocócicas/diagnóstico , Valor Preditivo dos Testes , Streptococcus pneumoniae/isolamento & purificação , Resultado do Tratamento , Procedimentos Cirúrgicos Vasculares
4.
PLoS One ; 14(2): e0207138, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30742636

RESUMO

BACKGROUND: Clostridium difficile (CD) is the leading cause of infectious health-care associated diarrhea. However, little is known regarding CD carriage and transmission amongst asymptomatic colonizers. We evaluated carriage, characterized strains and examined epidemiologic linkages in asymptomatic colonized CD patients. METHODS: Rectal swabs from asymptomatic patients admitted to the general medicine ward from April 1-June 30 2012 were collected. PCR-confirmed CD colonies were ribotyped and characterized by Modified-Multi Locus Variable Number Tandem Repeat Analysis (MMLVA). RESULTS: 1549-swabs were collected from 474-patients. Overall, 50/474(10.6%) were CD PCR-positive, 24/50 were colonized at admission, while 26/50 were first identified > = 72 hours after admission. Amongst the 50 CD PCR-positive patients, 90% were asymptomatically colonized and 80% of individuals carried toxigenic CD-strains, including ribotype-027 (5/45:11%). MMLVA revealed five-clusters involving 15-patients harboring toxigenic (4/5) and non-toxigenic CD strains (1/5). In two clusters, patients were CD positive on admission while in the other three clusters involving 10 patients, we observed CD transmission from asymptomatically colonized patients to 8 previously CD-negative patients. CONCLUSIONS: We identified increasing rates of colonization during admission to medical wards. MMLVA typing effectively discriminated between strains and suggests that 20% of patients with CD colonization acquired their strain(s) from asymptomatically colonized individuals in hospital.


Assuntos
Portador Sadio/microbiologia , Infecções por Clostridium/microbiologia , Clostridium difficile/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Infecção Hospitalar/microbiologia , Diarreia/microbiologia , Fezes/microbiologia , Feminino , Hospitalização , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Reto/microbiologia , Ribotipagem/métodos , Centros de Atenção Terciária , Adulto Jovem
5.
J Infect Chemother ; 25(4): 262-266, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30642771

RESUMO

In this study, we investigated all Clostridioides difficile strains isolated from stool samples in Nagasaki University Hospital between January 2012 and December 2014. Toxin genes (tcdA, tcdB and cdtA/cdtB) were analyzed for multiplex PCR in a total of 213 strains. In the toxin gene-positive strain, PCR ribotyping was conducted using capillary gel electrophoresis-based PCR and the Webribo database. Patients' backgrounds were analyzed by departments, disorders, antimicrobials, and clinical dates. The positive rates of tcdA, tcdB, and cdtA/cdtB genes were 62.9%, 63.4%, and 2.8%, respectively. The most frequent PCR ribotype was 047 (14.1%), followed by 014/0 (11.1%) and 002/0 (8.2%). In univariate analysis, the risk factors for the detection of toxin gene-positive strains in patients were older age (p = 0.0036), over ≥ 65 years old (p = 0.0175), the patients hospitalized at Department of Digestive Surgery (P = 0.0059), higher CRP level (P = 0.0395), and lower albumin level (p = 0.0014). In the multivariate analysis, the risk factor for detection of toxin gene-positive strains was the patients hospitalized at Department of Digestive Surgery (OR; 4.62, 95% CI; 1.18-18.0, p = 0.0274). In this study, the percentage of toxin gene-positive and cdtA/cdtB gene-positive strains was almost the same as that reported in previous studies, but the ribotype was different. In addition, we revealed that the risk factor associated with the detection of toxin gene-positive strains was the patients hospitalized at Department of digestive surgery.


Assuntos
Infecções por Clostridium/epidemiologia , Clostridium difficile/genética , Ribotipagem/métodos , ADP Ribose Transferases/genética , ADP Ribose Transferases/isolamento & purificação , Adolescente , Adulto , Fatores Etários , Idoso , Antibacterianos/uso terapêutico , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Infecções por Clostridium/tratamento farmacológico , Infecções por Clostridium/microbiologia , Clostridium difficile/isolamento & purificação , Fezes/microbiologia , Feminino , Hospitais Universitários/estatística & dados numéricos , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase/métodos , Fatores de Risco , Adulto Jovem
6.
Folia Microbiol (Praha) ; 64(1): 49-54, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29971567

RESUMO

This study aimed to implement a toxigenic culture as an optional third diagnostic step for glutamate dehydrogenase (GDH)-positive and toxin A/B-negative diarrheal stool samples into a diagnostic algorithm for Clostridioides (Clostridium) difficile infection (CDI), and to characterise C. difficile isolates for epidemiological purposes. During the 5-month study, 481 diarrhoeal stool samples from three Slovak hospitals were investigated and 66 non-duplicated GDH-positive stool samples were found. Of them, 36 were also toxin A/B-positive. Twenty-three GDH-positive and toxin A/B-negative stool samples were shown subsequently to be positive following toxigenic culture (TC). Molecular characterisation of C. difficile isolates showed the predominance of PCR ribotype (RT) 001 (n = 37, 56.1%) and the occurrence of RT 176 (n = 3, 4.5%). C. difficile RT 001 isolates clustered to eight clonal complexes (CCs) using multiple-locus variable-number tandem repeats analysis (MLVA). Interestingly, one third of RT 001 isolates clustering in these CCs were cultured from toxin A/B-negative stool samples. Our observations highlight the need of use multiple step diagnostic algorithm in CDI diagnosis in order to detect all CDI cases and to avoid the spread of C. difficile in healthcare settings.


Assuntos
Infecções por Clostridium/microbiologia , Clostridium difficile/classificação , Clostridium difficile/isolamento & purificação , Diarreia/diagnóstico , Tipagem Molecular , Reação em Cadeia da Polimerase , Ribotipagem/métodos , Algoritmos , Proteínas de Bactérias/análise , Infecções por Clostridium/diagnóstico , Infecções por Clostridium/epidemiologia , Análise por Conglomerados , Diarreia/epidemiologia , Diarreia/microbiologia , Enterotoxinas/análise , Enterotoxinas/deficiência , Fezes/microbiologia , Glutamato Desidrogenase/análise , Hospitais , Humanos , Repetições Minissatélites/genética , Técnicas de Diagnóstico Molecular , Tipagem de Sequências Multilocus , Eslováquia/epidemiologia
7.
Sci Rep ; 8(1): 15965, 2018 10 29.
Artigo em Inglês | MEDLINE | ID: mdl-30374098

RESUMO

In order to improve the diagnosis of pathogenic bacteria in cerebrospinal fluid (CSF) with purulent meningitis, we developed a DNA microarray technique for simultaneous detection and identification of seven target bacterium. DNA were extracted from 24 CSF samples with purulent meningitis (or suspected purulent meningitis). The specific genes of each pathogen were chosen as the amplification target, performed the polymerase chain reaction (PCR), labeled with a fluorescence dye, and hybridized to the oligonucleotide probes on the microarray. There is no significant cross-hybridization fluorescent signal occurred in untargeted bacteria. There were 87.5% (21/24) positive results in DNA microarray compared with the 58.3% (14/24) of the CSF culture test. Of which 58.3% (14/24) of the patients with culture-confirmed purulent meningitis, 37.5% (9/24) patients who were not confirmed by culture test but were demonstrated by the clinical diagnosis and DNA microarray. Multiple bacterial infections were detected in 5 cases by the microarray. In addition, the number of gene copies was carried out to determine the sensitivity of this technique, which was shown to be 3.5 × 101 copies/µL. The results revealed that the microarray technique which target pathogens of the CSF specimen is better specificity, accuracy, and sensitivity than traditional culture method. The microarray method is an effective tool for rapidly detecting more target pathogens and identifying the subtypes of strains which can eliminate the impact of the different individuals with purulent meningitis for prompt diagnosis and treatment.


Assuntos
DNA Bacteriano/líquido cefalorraquidiano , Meningites Bacterianas/microbiologia , Análise de Sequência com Séries de Oligonucleotídeos , Análise Serial de Tecidos/métodos , Adolescente , Adulto , Idoso , Técnicas Bacteriológicas , Líquido Cefalorraquidiano/microbiologia , Criança , Pré-Escolar , DNA Bacteriano/isolamento & purificação , Feminino , Genes Bacterianos , Humanos , Lactente , Masculino , Meningites Bacterianas/líquido cefalorraquidiano , Meningites Bacterianas/diagnóstico , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase/métodos , Ribotipagem/métodos , Sensibilidade e Especificidade , Adulto Jovem
8.
Sci Rep ; 8(1): 13951, 2018 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-30224751

RESUMO

The population structure of Clostridium difficile currently comprises eight major genomic clades. For the highly divergent C-I clade, only two toxigenic strains have been reported, which lack the tcdA and tcdC genes and carry a complete locus for the binary toxin (CDT) next to an atypical TcdB monotoxin pathogenicity locus (PaLoc). As part of a routine surveillance of C. difficile in stool samples from diarrheic human patients, we discovered three isolates that consistently gave negative results in a PCR-based screening for tcdC. Through phenotypic assays, whole-genome sequencing, experiments in cell cultures, and infection biomodels we show that these three isolates (i) escape common laboratory diagnostic procedures, (ii) represent new ribotypes, PFGE-types, and sequence types within the Clade C-I, (iii) carry chromosomal or plasmidal TcdBs that induce classical or variant cytopathic effects (CPE), and (iv) cause different levels of cytotoxicity and hamster mortality rates. These results show that new strains of C. difficile can be detected by more refined techniques and raise questions on the origin, evolution, and distribution of the toxin loci of C. difficile and the mechanisms by which this emerging pathogen causes disease.


Assuntos
Toxinas Bacterianas/genética , Cromossomos Bacterianos/genética , Clostridium difficile/genética , Clostridium difficile/patogenicidade , Virulência/genética , Proteínas de Bactérias/genética , Linhagem Celular Tumoral , Testes Diagnósticos de Rotina/métodos , Enterotoxinas/genética , Células HeLa , Humanos , Filogenia , Ribotipagem/métodos , Sequenciamento Completo do Genoma/métodos
10.
PLoS One ; 13(7): e0201539, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30048534

RESUMO

BACKGROUND: Clostridium difficile infection (CDI) caused by ribotype 002 strain is associated with poor outcomes in Chinese patients. Fecal microbiota transplantation (FMT) is an effective but costly treatment for CDI. We aimed to examine potential cost-effectiveness of ribotype-guided FMT in Chinese patients with severe CDI. METHODS: A decision-analytic model was designed to simulate outcomes of ribotype 002-guided FMT versus vancomycin treatment in Chinese patients with severe CDI in the hospital setting. Outcome measures included mortality rate; direct medical cost; and quality-adjusted life year (QALY) loss for CDI. Sensitivity analysis was performed to examine robustness of base-case results. RESULTS: Comparing to vancomycin treatment, ribotype-guided FMT group reduced mortality (11.6% versus 17.1%), cost (USD8,807 versus USD9,790), and saved 0.472 QALYs in base-case analysis. One-way sensitivity analysis found the ribotype-guided FMT group to remain cost-effective when patient acceptance rate of FMT was >0.6% and ribotype 002 prevalence was >0.07%. In probabilistic sensitivity analysis, ribotype-guided FMT gained higher QALYs at 100% of simulations with mean QALY gain of 0.405 QALYs (95%CI: 0.400-0.410; p<0.001). The ribotype-guided group was less costly in 97.9% of time, and mean cost-saving was USA679 (95%CI: 670-688; p<0.001). CONCLUSIONS: In the present model, ribotype-guided FMT appears to be a potential option to save QALYs and cost when comparing with vancomycin. The cost-effectiveness of ribotype-guided FMT is subject to the patient acceptance to FMT and prevalence of ribotype 002.


Assuntos
Infecções por Clostridium/terapia , Clostridium difficile/genética , Enterocolite Pseudomembranosa/terapia , Transplante de Microbiota Fecal/economia , Transplante de Microbiota Fecal/métodos , Ribotipagem , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/uso terapêutico , Grupo com Ancestrais do Continente Asiático , China/epidemiologia , Infecções por Clostridium/economia , Infecções por Clostridium/epidemiologia , Clostridium difficile/classificação , Análise Custo-Benefício , Técnicas de Apoio para a Decisão , Enterocolite Pseudomembranosa/economia , Enterocolite Pseudomembranosa/epidemiologia , Transplante de Microbiota Fecal/estatística & dados numéricos , Feminino , Frequência do Gene , Humanos , Masculino , Pessoa de Meia-Idade , Ribotipagem/economia , Ribotipagem/métodos , Índice de Gravidade de Doença , Taxa de Sobrevida , Resultado do Tratamento , Vancomicina/uso terapêutico
11.
Anaerobe ; 54: 1-7, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30009944

RESUMO

Restriction endonuclease analysis (REA) and PCR ribotyping are two typing systems that have been frequently utilized for molecular epidemiologic characterization of Clostridioides (Clostridium) difficile. To correlate typing data obtained from each method, we performed both REA and PCR ribotyping on a large and diverse set of historical and contemporary C. difficile infection clinical isolates. Eighty isolates were selected from each reference laboratory in the United States (Microbiology Reference Laboratory, Hines VA Medical Center) and United Kingdom (Clostridium difficile Network for England and Northern Ireland laboratory, University of Leeds). The 160 isolates were assigned to 82 unique ribotypes and 51 unique REA groups (116 unique REA types). In general, concordance between typing methods was good. Dendrogram analysis of PCR ribotype band patterns demonstrated close genetic relationships among strain types with discordant REA and ribotype assignments. While REA typing was more discriminatory, several REA types in this study were further discriminated by PCR ribotyping, indicating that discriminatory value of these typing methods may be strain dependent. These data will assist with molecular epidemiologic surveillance of strains identified by these two commonly used C. difficile typing systems.


Assuntos
Infecções por Clostridium/microbiologia , Clostridium difficile/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Mapeamento por Restrição/métodos , Ribotipagem/métodos , Clostridium difficile/classificação , Clostridium difficile/genética , Enzimas de Restrição do DNA/química , Humanos , Filogenia
12.
J Infect Chemother ; 24(8): 641-647, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29685855

RESUMO

We conducted a nationwide molecular epidemiological study of Clostridium difficile infection (CDI) in Japan investigated the correlation between the presence of binary toxin genes and CDI severity. This is the first report on molecular epidemiological analyses for CDI in multiple university hospitals in Japan, to our knowledge. We examined 124,484 hospitalized patients in 25 national and public university hospitals in Japan between December 2013 and March 2014, investigating antimicrobial susceptibilities and toxin-related genes for C. difficile isolates from stools. Epidemiological genetic typing was performed by PCR-ribotyping and repetitive sequence-based (rep)-PCR to examine the genetic similarities. The results detected toxin A-positive, toxin B-positive, binary toxin-negative (A+B+CDT-) detected from 135 isolates (80.8%) and toxin A-negative, toxin B-positive, binary toxin-negative (A- B+CDT-) in 23 (13.8%). Toxin A-positive, toxin B-positive, and binary toxin-positive (A+B+CDT+) were seen in 9 isolates (5.4%). Vancomycin (n = 81, 37.7%) or metronidazole (n = 88, 40.9%) therapies were undertaken in analyzed cases. Ribotypes detected from isolates were 017/subgroup 1, 070, 078, 126, 176, 449, 475/subgroup 1, 499, 451, 566 and newtypes. Rep-PCR classified 167 isolates into 28 cluster groups including 2-15 isolates. In addition, 2 pairs of strains isolated from different institutions belonged to the same clusters. Seven out of 9 (77.8%) of the patients with binary toxin producing strains had "mild to moderate" outcome in evaluated symptoms. In conclusion, we found that binary toxin did not show regional specificity and had no relevance to severity of CDI.


Assuntos
Antibacterianos/uso terapêutico , Infecções por Clostridium/epidemiologia , Clostridium difficile/genética , Hospitais Universitários/estatística & dados numéricos , ADP Ribose Transferases/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas de Bactérias/genética , Infecções por Clostridium/tratamento farmacológico , Infecções por Clostridium/microbiologia , Clostridium difficile/efeitos dos fármacos , Clostridium difficile/isolamento & purificação , Monitoramento Epidemiológico , Fezes/microbiologia , Feminino , Humanos , Concentração Inibidora 50 , Japão/epidemiologia , Masculino , Metronidazol/uso terapêutico , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Epidemiologia Molecular , Reação em Cadeia da Polimerase , Ribotipagem/métodos , Índice de Gravidade de Doença , Vancomicina/farmacologia , Vancomicina/uso terapêutico , Adulto Jovem
14.
Adv Exp Med Biol ; 1050: 45-58, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29383663

RESUMO

Clostridium difficile is a major bacterial cause of post-antibiotic diarrhoea. The epidemiology of C. difficile infections (CDI) has dramatically changed since the early 2000s, with an increasing incidence and severity across Europe. This trend is partly due to the emergence and rapid worldwide spread of the hypervirulent and epidemic PCR ribotype 027. Profiles of patients with CDI have also evolved, with description of community-acquired (CA) infections in patients with no traditional risk factors for CDI. However, recent epidemiological studies indicated that some European countries have successfully controlled the dissemination of the 027 clone whereas other countries recently reported the emergence of other virulent or unusual strains. The aims of this review are to summarize the current European CDI epidemiology and to describe the new virulent C. difficile strains circulating in Europe, as well as other potential emerging strains described elsewhere. Standardized typing methods and surveillance programmes are mandatory for a better understanding and monitoring of CDI in Europe.


Assuntos
Clostridium difficile/genética , Clostridium difficile/patogenicidade , Ribotipagem/métodos , Infecções por Clostridium/epidemiologia , Infecções por Clostridium/microbiologia , Infecções Comunitárias Adquiridas/epidemiologia , Infecções Comunitárias Adquiridas/microbiologia , Europa (Continente)/epidemiologia , Humanos , Reação em Cadeia da Polimerase , Virulência
15.
Int J Med Microbiol ; 308(3): 358-363, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29478838

RESUMO

Clostridium (Clostridioides) difficile is the main cause for nosocomial diarrhoea in industrialised nations. Epidemiologic data on the pathogen's occurrence in other world regions are still scarce. In this context we characterized with phenotypic and molecular genetic methods C. difficile isolates stemming from hospitalised patients with diarrhoea in Lebanon. From 129 stool samples of symptomatic patients at a tertiary care University hospital in Lebanon, a total of 107 C. difficile strains were cultivated and underwent ribotyping, toxin gene detection and antibiotic resistance testing. Ribotype 014 (RT014, 16.8%) predominated, followed by RT002 (9.3%), RT106 (8.4%) and RT070 (6.5%). Binary toxin gene-positive isolates (RT023, RT078 and RT126) were rarely detected and RT027 was absent. Interestingly, within one isolate only the toxin A gene (tcdA) was detected. Multiple-locus variable-number tandem repeat analysis (MLVA) revealed strong strain diversity in most RTs. The isolates were sensitive to metronidazole and vancomycin, and only a small proportion of strains displayed resistance against moxifloxacin, rifampicin, and clarithromycin (5.6%, 1.9%, and 2.8%), respectively. The data indicate that the genetic strain composition of Lebanese strains differs markedly from the situation seen in Europe and North America. Especially the epidemic RTs seen in the latter regions were almost absent in Lebanon. Interestingly, most strains showed almost no resistance to commonly used antibiotics that are suspected to play a major role in the development of C. difficile infection, despite frequent use of these antibiotics in Lebanon. Thus, the role of antimicrobial resistance as a major driving force for infection development remains uncertain in this area.


Assuntos
Antibacterianos/farmacologia , Toxinas Bacterianas/genética , Infecções por Clostridium/microbiologia , Clostridium difficile/efeitos dos fármacos , Clostridium difficile/genética , Farmacorresistência Bacteriana Múltipla , Toxinas Bacterianas/isolamento & purificação , Infecções por Clostridium/epidemiologia , Clostridium difficile/isolamento & purificação , Clostridium difficile/patogenicidade , Enterocolite Pseudomembranosa/epidemiologia , Enterocolite Pseudomembranosa/microbiologia , Enterotoxinas/genética , Fezes/microbiologia , Feminino , Fluoroquinolonas/farmacologia , Humanos , Líbano/epidemiologia , Masculino , Metronidazol/farmacologia , Testes de Sensibilidade Microbiana , Moxifloxacina , Tipagem de Sequências Multilocus/métodos , Fenótipo , Ribotipagem/métodos , Vancomicina/farmacologia
16.
Microb Drug Resist ; 24(4): 386-392, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29485939

RESUMO

OBJECTIVE: To assess drug susceptibility and characterize Clostridium difficile ribotypes in isolates from two tertiary-care hospitals in Mexico. METHODS: Isolates were evaluated for genotyping, antimicrobial susceptibility testing and detection of mutations associated with drug resistance. PCR ribotyping was performed using a combination of gel-based and capillary electrophoresis-based approaches. RESULTS: MIC50 and MIC90 were ≥128 mg/L for ciprofloxacin, erythromycin, clindamycin, and rifampicin. There was no reduced susceptibility to metronidazole or tetracycline; however, reduced susceptibility to vancomycin (≥4 mg/L) and fidaxomicin (≥2 mg/L) was detected in 50 (40.3%) and 4 (3.2%) isolates, respectively. Furthermore, the rpoB Arg505Lys mutation was more frequently detected in isolates with high minimum inhibitory concentration (MIC) to rifampicin (≥32 mg/L) (OR = 52.5; 95% CI = 5.17-532.6; p < 0.000). Of the 124 C. difficile isolates recovered, 84 (66.7%) were of ribotype 027, 18 (14.5%) of ribotype 001, and the remainder were other ribotypes (353, 255, 220, 208, 176, 106, 076, 020, 019, 017, 014, 012, 003, and 002). CONCLUSION: Ribotypes 027 and 001 were the most frequent C. difficile isolates recovered in this study, and demonstrated higher MICs. Furthermore, we found four isolates with reduced susceptibility to fidaxomicin, raising a concern since this drug is currently unavailable in Mexican Hospitals.


Assuntos
Clostridium difficile/genética , Clostridium difficile/isolamento & purificação , Farmacorresistência Bacteriana/genética , Antibacterianos/farmacologia , Infecções por Clostridium/tratamento farmacológico , Infecções por Clostridium/microbiologia , Clostridium difficile/efeitos dos fármacos , Farmacorresistência Bacteriana/efeitos dos fármacos , Humanos , México , Testes de Sensibilidade Microbiana/métodos , Ribotipagem/métodos , Centros de Atenção Terciária
17.
J Microbiol Methods ; 147: 50-55, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29486225

RESUMO

Clostridium difficile is the causative pathogen for antibiotic-related nosocomial diarrhea. For epidemiological study and identification of virulent clones, a new binary typing method was developed for C. difficile in this study. The usefulness of this newly developed optimized 10-loci binary typing method was compared with two widely used methods ribotyping and multilocus sequence typing (MLST) in 189 C. difficile samples. The binary typing, ribotyping and MLST typed the samples into 53 binary types (BTs), 26 ribotypes (RTs), and 33 MLST sequence types (STs), respectively. The typing ability of the binary method was better than that of either ribotyping or MLST expressed in Simpson Index (SI) at 0.937, 0.892 and 0.859, respectively. The ease of testing, portability and cost-effectiveness of the new binary typing would make it a useful typing alternative for outbreak investigations within healthcare facilities and epidemiological research.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Clostridium difficile/classificação , Clostridium difficile/isolamento & purificação , Tipagem de Sequências Multilocus/métodos , Ribotipagem/métodos , Proteínas de Bactérias/genética , Infecções por Clostridium/diagnóstico , Infecções por Clostridium/microbiologia , Clostridium difficile/genética , Clostridium difficile/patogenicidade , Simulação por Computador , DNA Bacteriano/análise , Loci Gênicos , Genótipo , Humanos , Epidemiologia Molecular/métodos , Tipagem Molecular/métodos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Fatores de Tempo
18.
J Microbiol Immunol Infect ; 51(3): 344-351, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28583353

RESUMO

PURPOSE: To characterise and compare twenty-eight Finnish Clostridium difficile RT027-like isolates, selected based on the presence of 18 bp deletion in the tcdC gene and toxin gene profile (A, B, binary), with eleven RT027 isolates from different Finnish geographical areas and time periods. METHODS: Twenty-eight C. difficile RT027-like isolates and 11 RT027 comparative strains were characterised by capillary-electrophoresis (CE) ribotyping, multi-locus variable tandem-repeats analysis (MLVA), multi-locus sequence typing (MLST), and sequencing of tcdC and gyrA gene fragments. Susceptibility to moxifloxacin was determined by E-test. RESULTS: Of 28 RT027-like isolates, seven RTs (016, 034, 075, 080, 153, 176 and 328), three WEBRIBO types (411, 475, AI-78) and three new profiles (F1-F3) were identified. MLVA revealed six clonal complexes (RTs 016, 027, 176 and F3). MLST showed eleven sequence types (1, 41, 47, 67, 95, 191,192, 223, 229, 264 and new ST). Twenty-two isolates (RTs 016, 080, 176, 328, F1, F2, F3 and WRTAI-78) carried Δ117 in the tcdC gene. Isolates of RTs 016, 027 and 176 were moxifloxacin resistant and harboured Thr82Ile in the GyrA. CONCLUSION: Our results show a high diversity within 28 Finnish RT027-like C. difficile isolates, with twelve CE-ribotyping profiles and eleven STs. MLVA revealed the regional spread of RTs 016, 027, 176 and F3. The presence of Δ117 in the tcdC gene in eight non-027 RTs highlights the importance of careful interpretation of the results from molecular systems targeting this site in the genome of C. difficile and the need of strain typing for epidemiological purposes.


Assuntos
Técnicas de Tipagem Bacteriana , Clostridium difficile/genética , Clostridium difficile/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Ribotipagem/métodos , ADP Ribose Transferases/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Toxinas Bacterianas/análise , Toxinas Bacterianas/genética , Sequência de Bases , Infecções por Clostridium/epidemiologia , Infecções por Clostridium/microbiologia , Clostridium difficile/classificação , Clostridium difficile/efeitos dos fármacos , DNA Girase/genética , DNA Bacteriano/análise , Farmacorresistência Bacteriana , Enterotoxinas/genética , Feminino , Finlândia , Fluoroquinolonas/farmacologia , Genótipo , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Repetições Minissatélites/genética , Epidemiologia Molecular , Moxifloxacina , Tipagem de Sequências Multilocus/métodos , Proteínas Repressoras/genética , Adulto Jovem
19.
Food Res Int ; 102: 68-76, 2017 12.
Artigo em Inglês | MEDLINE | ID: mdl-29196000

RESUMO

The pit mud (PM) in fermentation cellar is a complex ecosystem that hosts diverse microbial communities that contribute to the production of Chinese strong-flavor Baijiu (CSFB). However, the microbial ecology of PM, particularly the extent of their phylogenetic novelty remains poorly understood. Here we conducted Illumina MiSeq sequencing to explore the diversity and novelty patterns of PM bacterial communities from Luzhou Laojiao cellars in use for 40 and 400years. High diversity indices were found in the PM with 16 phyla and 105 genera. Interestingly, the compositions of dominant genera of the PM were significantly different than that reported previously for PM sampled from other geographic sites, suggesting greater microbial diversity of PM. The dominant genus of Caproiciproducens, a caproic acid-producing bacterium, is the first reported for Chinese Baijiu production. Our results demonstrate that the PM hosts a large number of novel taxa, with 26% of the total OTUs (operational taxonomic units) distant to cultured counterparts. The class Clostridia within Firmicutes presented the highest proportion of novel OTUs. Most novel OTUs were initially isolated from diverse environments, the most abundant of which came from Chinese Baijiu brewing ecosystems, highlighting the huge culturing gap within the PM, but at the same time suggesting the importance of these OTUs in CSFB production. The data presented in this study significantly increases the number of bacteria known to be associated with CSFB production and should help guide the future exploration of microbial resources for biotechnological applications.


Assuntos
Bebidas Alcoólicas/microbiologia , Bactérias/genética , DNA Bacteriano/genética , Fermentação , Microbiologia de Alimentos/métodos , Sequenciamento de Nucleotídeos em Larga Escala , Filogenia , Ribotipagem/métodos , Microbiologia do Solo , Bactérias/classificação , Bactérias/crescimento & desenvolvimento , Bactérias/isolamento & purificação , China , DNA Bacteriano/classificação , DNA Bacteriano/isolamento & purificação , Genótipo , Fenótipo
20.
PLoS One ; 12(12): e0189183, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29287077

RESUMO

BACKGROUND: It has been suggested that the high incidence of ribotype 078 Clostridium difficile infections (CDI) in the Netherlands is related to pig farming. METHODS: We used data of hospitalised CDI patients (>2yrs of age) diagnosed between May 2009 and May 2015 in 26 hospitals participating in a national sentinel surveillance. We compared clinical and geographical characteristics of 078 CDI to other CDI. We investigated the association between 078 CDI incidence and four indicators of pig farming (piglet, pig, piglet farm and pig farm density) by mixed-effects Poisson regression. We used a space-time permutation model to search for community-onset 078 CDI clusters (using SaTScan). RESULTS: A total of 4,691 CDI were identified. Ribotype 078 was isolated in 493 of 3,756 patients (13.1%) including a typing result. These patients had slightly higher community-onset disease and a 35% increase of 30-day mortality compared to non-078 CDI patients. The pooled overall and 078 incidence rates were 2.82 (95% CI, 2.42-3.29) and 0.26 (95% CI, 0.21-0.31) CDI per 10,000 patients-days respectively. Hospital 078 CDI incidence was not associated with provincial pig (IRR, 0.98; 95% CI, 0.89-1.08), piglet (IRR, 0.95; 95% CI, 0.75-1.19), pig farm (IRR, 1.08; 95% CI, 0.84-1.39), or piglet farm density (IRR, 1.00; 95% CI, 0.56-1.79). No clusters of community-onset ribotype 078 CDI were found. CONCLUSIONS: Our results do not indicate that the ribotype 078 CDI incidence in hospitals is related to pig (farm) or piglet (farm) density. However, transmission beyond provincial borders or in non-hospitalised patients cannot be excluded.


Assuntos
Agricultura , Infecções por Clostridium/epidemiologia , Clostridium difficile/patogenicidade , Ribotipagem/métodos , Vigilância de Evento Sentinela , Idoso , Animais , Infecções por Clostridium/microbiologia , Infecções por Clostridium/transmissão , Análise por Conglomerados , Feminino , Humanos , Incidência , Masculino , Países Baixos/epidemiologia , Distribuição de Poisson , Suínos
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