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1.
Avian Pathol ; 50(1): 31-40, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-32990455

RESUMEN

A total of 62 isolates of Riemerella-like organisms, originally isolated from Australian poultry (10 from chickens, 46 from ducks, five from unknown hosts and one vaccine strain), were included in this study. On the basis of two published polymerase chain reaction (PCR) assays that are reported to be specific for Riemerella anatipestifer, 51 of the isolates were identified as R. anatipestifer. Forty-six of these isolates had a detailed history and were sourced from ducks, while five were of unknown origin. The 11 remaining isolates failed to yield a positive reaction in either PCR with 10 originating from chickens and one from a duck. Amplification and sequencing of the 16S rRNA gene of these isolates identified the duck isolate as Moraxella lacunta. Phylogenetic analysis of the 10 chicken isolates identified one as R. columbina and the remaining nine isolates as Riemerella-like taxon 2. The 51 Australian R. anatipestifer isolates were assigned by gel diffusion test to serovars 1 (26 isolates), 6 (seven isolates), 8 (five isolates), 9 (two isolates), 13 (one isolate) and 14 (one isolate) while nine isolates gave no reaction to any antiserum. A commercial system was used to perform DNA fingerprinting using rep-PCR analysis, which revealed different clusters with a lack of a clear relationship between the clusters and the serovars.


Asunto(s)
Pollos/virología , Patos/virología , Infecciones por Flavobacteriaceae/veterinaria , Enfermedades de las Aves de Corral/virología , Riemerella/inmunología , Animales , Australia , Infecciones por Flavobacteriaceae/virología , Filogenia , Reacción en Cadena de la Polimerasa/veterinaria , ARN Ribosómico 16S/genética , Riemerella/clasificación , Riemerella/genética , Serogrupo
2.
Niger J Clin Pract ; 23(8): 1155-1162, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32788495

RESUMEN

OBJECTIVE: The blaOXA resistance genes and ISAba1 were examined in 70 samples from lower respiratory tract of hospitalized patients. MATERIALS AND METHODS: Of the 67 isolates obtained, almost half (46.3%) of them were from endotracheal aspirate, and most were collected from the intensive care units of the reanimation (37.3%) and internal medicine (32.8%) units. RESULTS: Three samples from the internal medicine intensive care unit had positive cultures. Of the multidrug resistant (MDR) samples, 70 isolates (>50%) were moderately sensitive, while fewer (10%) were resistant to tigecycline. In contrast, 100% were sensitive to colistin. All strains were found to be positive for blaOXA-23-like and blaOXA-51-like genes, whereas no blaOXA-40-like and blaOXA-58-like genes were detected. The ISAba1 positivity rate was 90.0%. Pattern 5 was mainly identified among the 22 different patterns. Of note, 50% of Pattern 5 was found in the patients of the internal medicine intensive care unit, and a third was associated with ventilator-associated pneumonia. Importantly, the internal medicine unit's equipment was found to be culture positive. CONCLUSION: Findings obtained from this study suggest that isolates can easily spread through the hospital via isolate cross-contamination caused by health personnel. These contaminating isolates may be able to maintain their presence within the hospital for a long time.


Asunto(s)
Infecciones por Acinetobacter/tratamiento farmacológico , Acinetobacter baumannii/genética , Acinetobacter baumannii/aislamiento & purificación , Antibacterianos/farmacología , Farmacorresistencia Bacteriana Múltiple/genética , beta-Lactamasas/genética , Infecciones por Acinetobacter/microbiología , Acinetobacter baumannii/efectos de los fármacos , Adulto , Anciano , Anciano de 80 o más Años , Proteínas Bacterianas/genética , Colistina/farmacología , ADN Bacteriano/genética , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Femenino , Genes Bacterianos , Humanos , Unidades de Cuidados Intensivos , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Tipificación Molecular , Reacción en Cadena de la Polimerasa Multiplex , Centros de Atención Terciaria
3.
J Clin Microbiol ; 56(10)2018 10.
Artículo en Inglés | MEDLINE | ID: mdl-30068536

RESUMEN

Streptococcus zooepidemicus is an emerging and opportunistic zoonotic pathogen which plays an important role in the development of severe and life-threatening diseases and is potentially capable of triggering large glomerulonephritis outbreaks. Between December 2012 and February 2013, 175 cases of glomerulonephritis were confirmed in the town of Monte Santo de Minas, MG, Brazil. During the outbreak, 19 isolates of S. zooepidemicus were recovered, 1 from ice cream, 2 from the oropharynx of food handlers, and 16 from patients affected by acute poststreptococcal glomerulonephritis (APSGN). All S. zooepidemicus isolates involved in the outbreak amplified the same sequence of the hypervariable region of the SzP protein (SzPHV5) and presented indistinguishable banding patterns with high similarity (>99%) to each other by the repetitive element sequence-based PCR (rep-PCR) technique. Inspection programs on the milk supply chain should be strengthened and continuously encouraged so that the health of consumers is preserved.


Asunto(s)
Brotes de Enfermedades , Glomerulonefritis/epidemiología , Glomerulonefritis/microbiología , Infecciones Estreptocócicas/epidemiología , Infecciones Estreptocócicas/microbiología , Streptococcus equi/aislamiento & purificación , Adulto , Animales , Proteínas Bacterianas/genética , Técnicas Bacteriológicas , Brasil/epidemiología , ADN Bacteriano/genética , Femenino , Microbiología de Alimentos , Humanos , Masculino , Leche/microbiología , Reacción en Cadena de la Polimerasa , Infecciones Estreptocócicas/transmisión , Streptococcus equi/clasificación , Streptococcus equi/genética
4.
Mycoses ; 60(5): 348-354, 2017 May.
Artículo en Inglés | MEDLINE | ID: mdl-28220547

RESUMEN

Dermatophyte species, isolation and identification in clinical samples are still difficult and take a long time. The identification and molecular epidemiology of dermatophytes commonly isolated in a clinical laboratory in Turkey by repetitive sequence-based PCR (rep-PCR) were assessed by comparing the results with those of reference identification. A total of 44 dermatophytes isolated from various clinical specimens of 20 patients with superficial mycoses in Kayseri and 24 patients in Hatay were studied. The identification of dermatophyte isolates was based on the reference identification and rep-PCR using the DiversiLab System (BioMerieux). The genotyping of dermatophyte isolates from different patients was determined by rep-PCR. In the identification of dermatophyte isolates, agreement between rep-PCR and conventional methods was 87.8 % ( 36 of 41). The dermatophyte strains belonged to four clones (A -D) which were determined by the use of rep-PCR. The dermatophyte strains in Clone B, D showed identical patterns with respect to the region. In conclusion, rep-PCR appears to be useful for evaluation of the identification and clonal relationships between Trichophyton rubrum species complex and Trichophyton mentagrophytes species complex isolates. The similarity and diversity of these isolates may be assessed according to different regions by rep-PCR.


Asunto(s)
Arthrodermataceae/genética , Dermatoglifia del ADN/métodos , ADN de Hongos/genética , Dermatomicosis/epidemiología , Genotipo , Trichophyton/genética , Adolescente , Adulto , Anciano , Arthrodermataceae/clasificación , Arthrodermataceae/aislamiento & purificación , Niño , Dermatomicosis/diagnóstico , Dermatomicosis/microbiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa/métodos , Análisis de Secuencia de ADN , Trichophyton/clasificación , Turquía/epidemiología , Adulto Joven
5.
Anaerobe ; 47: 73-78, 2017 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-28454760

RESUMEN

The recognition of the pathogenicity of Cutibacterium acnes in implant-associated infection is not always obvious. In this paper, we aimed to distinguish pathogenic and non-pathogenic C. acnes isolates. To reach this goal, we investigated the clonal complex (CC) of a large collection of C. acnes clinical isolates through Multi-Locus Sequence Typing (MLST), we established a Caenorhabditis elegans model to assess C. acnes virulence and we investigated the presence of virulence factors in our collection. Ours results showed that CC36 and CC53 C. acnes isolates were more frequently observed in prosthetic joint infections (PJI) than CC18 and CC28 C. acnes isolates (p = 0.021). The C. elegans model developed here showed two distinct virulence groups of C. acnes (p < 0.05). These groups were not correlated to CC or clinical origin. Whole genome sequencing allowed us to identify a putative gene linked to low virulent strains. In conclusion, MLST remains a good method to screen pathogenic C. acnes isolates according to their clinical context but mechanisms of C. acnes virulence need to be assess thought transcriptomic analysis to investigate regulatory process.


Asunto(s)
Infecciones por Bacterias Grampositivas/microbiología , Propionibacterium acnes/fisiología , Propionibacterium acnes/patogenicidad , Infecciones Relacionadas con Prótesis/microbiología , Tropismo Viral , Animales , Caenorhabditis elegans/microbiología , Caenorhabditis elegans/fisiología , Modelos Animales de Enfermedad , Humanos , Tipificación de Secuencias Multilocus , Propionibacterium acnes/clasificación , Propionibacterium acnes/genética , Análisis de Supervivencia , Virulencia , Factores de Virulencia/análisis , Factores de Virulencia/genética , Secuenciación Completa del Genoma
6.
J Infect Chemother ; 22(6): 407-13, 2016 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-27107736

RESUMEN

Pneumococcal Molecular Epidemiology Network (PMEN) clones are representatives of worldwide-spreading pathogens. DiversiLab system, a repetitive PCR system, has been proposed as a less labor-and time-intensive genotyping platform alternative to conventional methods. However, the utility and analysis parameters of DiversiLab for identifying worldwide lineages was not established. To evaluate and optimize the performance of DiversiLab for identifying worldwide pneumococcal lineages, we examined 245 consecutive isolates of clinical Streptococcus pneumoniae from all age-group patients at a teaching hospital in Japan. The capsular swelling reaction of all isolates yielded 24 different serotypes. Intensive visual observation (VO) of DiversiLab band pattern difference divided all isolates into 73 clusters. Multilocus sequence typing (MLST) of representative 73 isolates from each VO cluster yielded 51 different STs. Among them, PMEN-related lineages accounted for 63% (46/73). Although the serotype of PMEN-related isolates was identical to that of the original PMEN clone in 70% (32/46), CC156-related PMEN lineages, namely Greece(6B)-22 and Colombia(23F)-26, harbored various capsular types discordant to the original PMEN clones. Regarding automated analysis, genotyping by extended Jaccard (XJ) with a 75% similarity index cutoff (SIC) showed the highest correlation with serotyping (adjusted Rand's coefficient, 0.528). Elevating the SIC for XJ to 85% increased the discriminatory power sufficient for distinguishing two major PMEN-related isolates of Taiwan(19F)-14 and Netherlands(3)-31. These results demonstrated a potential utility of DiversiLab for identifying worldwide lineage of pneumococcus. An optimized parameters of automated analysis should be useful especially for comparison for reference strains by "identification" function of DiversiLab.


Asunto(s)
Tipificación de Secuencias Multilocus , Reacción en Cadena de la Polimerasa , Streptococcus pneumoniae/clasificación , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Genotipo , Hospitales de Enseñanza , Humanos , Lactante , Japón , Persona de Mediana Edad , Tipificación de Secuencias Multilocus/métodos , Reacción en Cadena de la Polimerasa/métodos , Serotipificación , Streptococcus pneumoniae/efectos de los fármacos , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/aislamiento & purificación , Adulto Joven
7.
Mycoses ; 58(3): 149-59, 2015 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-25591136

RESUMEN

Two strains of Cryptococcus neoformans (PU 66 and PU112) were simultaneously isolated from a patient with systemic lupus erythematosus. We aimed to trace the source of the mixed infections. Multi-locus sequence typing (MLST) and the DiversiLab system analyses were performed on the 2 clinical and 23 environmental C. neoformans from pigeon droppings, 11 from the home (H1) the patient visited, 12 from another home (H2) as control. All the strains were uniformly genotyped as C. neoformans var. grubii VNI. Clinical strain PU66 and all the H1 isolates had the same sequence type (ST) - ST5, while for PU112 a new ST was observed - ST265. However, there was only one single base of 7 MLST loci difference between PU66 and PU112. Sequence types of the H2 strains were ST31 and ST297. DiversiLab analysis showed that strain similarity between the two clinical strains was 96.7%. In relation to environmental samples, the highest strain similarity (99.3%) was observed for PU66 and PU70 (H1). However, none of the environmental isolates had similarity over 98.6% comparing to PU112. One source of the mixed infections has been detected, but another needs further investigation.


Asunto(s)
Criptococosis/complicaciones , Criptococosis/microbiología , Cryptococcus neoformans/aislamiento & purificación , Lupus Eritematoso Sistémico/complicaciones , Adulto , Animales , Coinfección , Columbidae/microbiología , Cryptococcus neoformans/clasificación , ADN de Hongos/genética , Femenino , Genotipo , Humanos , Tipificación de Secuencias Multilocus , Mutación , Técnicas de Tipificación Micológica , Filogenia
8.
J Clin Lab Anal ; 29(3): 250-3, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-24796534

RESUMEN

We evaluated the DiversiLab (DL) system with universal primers, a semiautomated repetitive extragenic palindromic sequence-based polymerase chain reaction (PCR) (rep-PCR) system, for the characterization of Helicobacter pylori in Japan. All 135 isolates from Japanese patients with gastric cancer (GC, n = 55) or non-GC (n = 80) were used and subjected to the drug susceptibility examinations (amoxicillin, AMPC; metronidazole, MNZ; and clarithromycin, CAM) by E-test. There were 28 MNZ-resistant (20.7%), 35 CAM-resistant (25.9%), and 16 MNZ/CAM-resistant (11.9%) isolates. DL rep-PCR fingerprinting analysis at the level of 95% similarity revealed five major groups (A-E) and the other including 45 isolates. The occupation rates of GC-derived isolates in groups B (54.2%) and E (58.8%) were higher than in the other groups: A (26.7%), C (28.6%), D (30.0%), and the other (40.0%). Relative higher occupation rates of drug resistants, such as MNZ-, CAM- and double MNZ/CAM-resistant isolates, were observed in groups B (45.8%), C (42.6%), and D (40%). Five of eight GC-derived isolates with MNZ/CAM resistance were significantly assigned to group B (P = 0.0312, χ(2) -test). These results suggest that the isolates classified in group B have a potential to contribute to the development of severe gastric disorders. The DL system, rapid and high sensitive technology, would be widely available in clinical laboratory for pathological and epidemiological analyses even in H. pylori.


Asunto(s)
Infecciones por Helicobacter/diagnóstico , Helicobacter pylori/genética , Helicobacter pylori/patogenicidad , Reacción en Cadena de la Polimerasa/métodos , Distribución de Chi-Cuadrado , Femenino , Infecciones por Helicobacter/complicaciones , Helicobacter pylori/aislamiento & purificación , Humanos , Japón , Masculino , Neoplasias Gástricas/complicaciones , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/microbiología
9.
J Antimicrob Chemother ; 69(9): 2383-7, 2014 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-24562615

RESUMEN

OBJECTIVES: We examined the molecular epidemiology of Acinetobacter baumannii clinical isolates from two cities (Tehran and Tabriz) of Iran. METHODS: DiversiLab repetitive extragenic palindromic PCR (rep-PCR), multilocus sequence typing and sequence group multiplex PCR were performed. The presence of resistance mechanisms including metallo-ß-lactamases, extended-spectrum ß-lactamases, OXA carbapenemases, aminoglycoside-modifying enzymes and RNA methylases was also investigated. RESULTS: DiversiLab rep-PCR identified 11 clusters and 11 singleton isolates. Twelve sequence types (STs), including six novel types, were identified. Sequence groups (SGs) 1-3 as well as five additional banding patterns were detected by multiplex PCR. A local outbreak in a general hospital in Tabriz with an SG1/ST2 profile was identified. Isolates of international clone II showed the highest prevalence and the most heterogeneous combination of resistance determinants. CONCLUSIONS: Several different multiresistant strains of A. baumannii were shown to circulate in Iran. The selection and spread of the SG1/ST2 clone might have been favoured by the acquisition of resistance genes in the absence of adequate infection control measures.


Asunto(s)
Infecciones por Acinetobacter/epidemiología , Acinetobacter baumannii/clasificación , Acinetobacter baumannii/genética , Farmacorresistencia Bacteriana Múltiple , Tipificación Molecular , Infecciones por Acinetobacter/microbiología , Acinetobacter baumannii/efectos de los fármacos , Acinetobacter baumannii/aislamiento & purificación , Ciudades/epidemiología , Análisis por Conglomerados , Variación Genética , Genotipo , Humanos , Irán/epidemiología , Epidemiología Molecular
10.
J Mycol Med ; 28(2): 320-326, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29580647

RESUMEN

The objective of our study was to assess the DiversiLab® automated repetitive sequence-based PCR (rep-PCR) system for typing C. albicans and C. glabrata isolates and to compare it with two proven and routinely used typing methods. A total of 39 isolates from 11 patients with candidaemia or tissue candidiasis (two to six isolates per patient) were analyzed with three typing methods: DiversiLab® rep-PCR, multilocus sequence typing and multilocus microsatellite typing. DiversiLab® rep-PCR results were consistent with those obtained using the two other typing methods for C. albicans, but not for C. glabrata. Thanks to its simplicity of use, rapidity, standardization and reproducibility, the DiversiLab® rep-PCR system is an interesting tool to investigate C. albicans infections.


Asunto(s)
Candida albicans/clasificación , Candida glabrata/clasificación , Reacción en Cadena de la Polimerasa/métodos , Secuencias Repetitivas de Ácidos Nucleicos , Automatización , Candida albicans/aislamiento & purificación , Candida glabrata/aislamiento & purificación , Candidiasis/diagnóstico , Candidiasis/microbiología , Genotipo , Humanos , Repeticiones de Microsatélite , Tipificación de Secuencias Multilocus , Técnicas de Tipificación Micológica , Reacción en Cadena de la Polimerasa/normas , Reproducibilidad de los Resultados
11.
J Hosp Infect ; 99(2): 208-217, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29203445

RESUMEN

BACKGROUND: The current increase in nosocomial infections caused by vancomycin-resistant enterococci (VRE) warrants improvement of detection methods and hygiene measures. Knowledge of the local epidemiology is important for monitoring compliance of medical personnel with hygiene measures. AIM: To evaluate semi-automated repetitive element palindromic polymerase chain reaction (rep-PCR) for rapid molecular typing of VRE. METHODS: Primary VRE isolates were collected during an observation period of one year and retrospectively typed by rep-PCR. Molecular typing was performed on isolates from two departments with elevated VRE rates and patients with increased risk for systemic VRE infections. Typing results were correlated with temporal and spatial information on patient moves, VRE laboratory results and multi-locus sequence typing (MLST). FINDINGS: Approximately 70% of VRE isolates within a department could be assigned to similarity clusters. Spread of VRE was limited to the individual departments. There was no evidence for spread of endemic VRE strains within the geographical catchment area of the hospital. Our results demonstrate the utility of rep-PCR typing on a department level. However, a Diversilab® threshold of ≥98% had to be applied to claim similarity, and suspected transmissions needed to be confirmed by vanA/B genotyping and compiled information on spatial and temporal patient contact. MLST verified the findings. CONCLUSION: Spread of predominantly detected vancomycin-resistant Enterococcus faecium was limited to the department level with no evidence for wider dissemination within the hospital. Well-standardized and validated (semi-)automated rep-PCR systems are useful for rapid detection of possible VRE transmission. However, suspected transmissions need to be confirmed by clinical and microbiological parameters.


Asunto(s)
Infección Hospitalaria/epidemiología , Enterococcus faecium/aislamiento & purificación , Infecciones por Bacterias Grampositivas/epidemiología , Epidemiología Molecular/métodos , Tipificación Molecular/métodos , Reacción en Cadena de la Polimerasa/métodos , Enterococos Resistentes a la Vancomicina/aislamiento & purificación , Infección Hospitalaria/microbiología , Infección Hospitalaria/transmisión , ADN Bacteriano/genética , Enterococcus faecium/clasificación , Enterococcus faecium/genética , Monitoreo Epidemiológico , Infecciones por Bacterias Grampositivas/microbiología , Infecciones por Bacterias Grampositivas/transmisión , Departamentos de Hospitales , Humanos , Epidemiología Molecular/normas , Tipificación Molecular/normas , Reacción en Cadena de la Polimerasa/normas , Secuencias Repetitivas de Ácidos Nucleicos , Estudios Retrospectivos , Análisis Espacio-Temporal , Enterococos Resistentes a la Vancomicina/clasificación , Enterococos Resistentes a la Vancomicina/genética
12.
Clin Microbiol Infect ; 22(3): 285.e9-17, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26686810

RESUMEN

During recent years the proportion of tinea capitis infections due to Microsporum audouinii has increased in both Belgium and other European countries. To better understand the emergence of this species, the Belgian National Reference Centre for dermatophytes launched an epidemiological survey on the main anthropophilic dermatophytes causing tinea capitis in Belgium and included the genomic characterization of M. audouinii isolates. In total, 116 strains of M. audouinii were confirmed and characterized by the DiversiLab(®) system (bioMérieux). Six genotypic variants were identified, among which one major group included 90 isolates and the reference strain. Another variant group (11 strains) was exclusively confined to a geographical region in south Belgium. Analysis of epidemiological characteristics of the infected population showed that the main age category was 5- to 9-year-old children with a sex ratio (male/female) of 1.97. Data concerning the geographic origin of the family revealed a majority of Belgian nationality (44.7%), suggesting that the infection originated in Belgium. Other nationalities were primarily African. At this time, no clear correlation has been established between one particular strain and a specific country of origin.


Asunto(s)
Dermatomicosis/epidemiología , Dermatomicosis/microbiología , Genotipo , Microsporum/clasificación , Microsporum/genética , Adolescente , Bélgica/epidemiología , Niño , Preescolar , ADN Espaciador Ribosómico , Femenino , Genes Fúngicos , Humanos , Lactante , Recién Nacido , Masculino , Tipificación Molecular , Vigilancia de la Población , Análisis de Secuencia de ADN , Adulto Joven
13.
Ann Lab Med ; 36(6): 536-41, 2016 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-27578506

RESUMEN

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) are important pathogens causing nosocomial infections in Korean hospitals. This study aimed to investigate the epidemiological and genetic diversity of clinical S. aureus isolates in healthcare settings from 2001 to 2008. METHODS: Samples and data were obtained from 986 individuals as part of the National Antimicrobial Surveillance Project, involving 10 regions nationwide. Molecular typing studies, including multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing were performed, and a representative clone of Korean MRSA was classified by combinational grouping using a DiversiLab (DL; bioMérieux, France) repetitive element polymerase chain reaction (rep-PCR) system. RESULTS: Nine Korean MRSA clones (KMRSA-1 to -9) were identified by analysis of genetic backgrounds and molecular characteristics. KMRSA-1 to -3, expressing clonal complex (CC) 5 (carrying SCCmec II), CC8 (carrying SCCmec III), and CC72 (carrying SCCmec IV) were spread nationwide. In contrast, KMRSA-6 was highly prevalent in Gyeongsangnam-do, and KMRSA-4 was highly prevalent in Jeollanam-do and Jeollabuk-do. CONCLUSIONS: Epidemic KMRSA clones were genetically similar to major clones identified from the USA, with the exception of KMRSA-2, which had the SCCmec III type. Our results provide important insights into the distribution and molecular genetics of MRSA strains in Korea and may aid in the monitoring of MRSA spread throughout the country.


Asunto(s)
Staphylococcus aureus Resistente a Meticilina/genética , Infecciones Estafilocócicas/epidemiología , Proteínas Bacterianas/genética , ADN Bacteriano/genética , ADN Bacteriano/metabolismo , Electroforesis en Gel de Campo Pulsado , Hospitales , Humanos , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Tipificación de Secuencias Multilocus , Reacción en Cadena de la Polimerasa Multiplex , Prevalencia , República de Corea/epidemiología , Infecciones Estafilocócicas/diagnóstico , Infecciones Estafilocócicas/microbiología
14.
Infect Dis (Lond) ; 48(8): 596-603, 2016 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-27171371

RESUMEN

BACKGROUND: Saprochaete capitata isolates have emerged as important nosocomial pathogens, among immunosuppressed or neutropenic patients, and a rare cause of nosocomial infection in the hematology-bone marrow unit (HBMU) and the intensive care unit (ICU). The purpose of this study was to molecular epidemiology and antifungal susceptibility of S. capitata (Blastoschizomyces capitatus) isolates causing nosocomial infection at Kayseri in Turkey. METHODS: During a period from 2012 to 2015, a total of 20 S. capitata strains were obtained from patients hospitalized at Erciyes University Hospital. The identification of S. capitata was performed by phenotypic and biochemical methods; this was confirmed by molecular methods by DNA sequencing analysis. Genotyping of S.capitata isolates from different patients was determined to by the repetitive sequence PCR (repPCR) using the DiversiLab System (BioMerieux). RESULTS: More than half of the patients with S. capitata infections were hospitalized in the hematology-oncology unit (60%). The patients mainly included those using intravascular devices (90%), and receiving parenteral antibiotics (85%); the mortality rate was 55%. The microbiological investigation failed to identify S. capitata in the hospital environment. All isolates were resistant to caspofungin (>32). However, the MIC90 values for voriconazole, amphotericin B, and fluconazole against all of the isolates were 0.125, 0.25, and 1µg/ml, respectively. The S. capitata strains belonged to five clones (A-E) which were determined by the use of rep-PCR and Clone C was found to be predominant. CONCLUSIONS: S. capitata isolates are an important cause of nosocomial infection in the HBMU and ICUs.


Asunto(s)
Antifúngicos/farmacología , Infección Hospitalaria/microbiología , Micosis/microbiología , Saccharomycetales/efectos de los fármacos , Saccharomycetales/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Infección Hospitalaria/epidemiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Epidemiología Molecular , Micosis/epidemiología , Reacción en Cadena de la Polimerasa , Estudios Retrospectivos , Turquía/epidemiología , Adulto Joven
15.
J Microbiol Methods ; 118: 81-4, 2015 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-26341611

RESUMEN

We evaluated and critically assessed the performance and discriminatory power of a rep-PCR based commercial test DiversiLab® Enterococcus kit (bioMerieux) for typing a set of 65 representative isolates of Enterococcus faecium/VRE and compared it to state-of-the-art typing techniques such as PFGE and MLST.


Asunto(s)
Enterococcus faecium/clasificación , Enterococcus faecium/genética , Infecciones por Bacterias Grampositivas/microbiología , Tipificación Molecular/métodos , Electroforesis en Gel de Campo Pulsado/métodos , Enterococcus faecium/aislamiento & purificación , Infecciones por Bacterias Grampositivas/epidemiología , Humanos , Epidemiología Molecular/métodos , Tipificación de Secuencias Multilocus/métodos , Reacción en Cadena de la Polimerasa/métodos
16.
Infect Genet Evol ; 26: 14-9, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24815727

RESUMEN

Enterococcus is a major cause of important nosocomial infections. Linezolid, the first member of an entirely new class of antibiotics (oxazolidinones), is effective against serious infections caused by Enterococcus. However, resistance to linezolid has been discovered throughout the world rapidly. From 2011 to 2013, nine linezolid-resistant E. faecalis isolates were collected and the possible mechanisms of linezolid resistance, including mutations in domain V of 23S rRNA genes and in ribosomal proteins L3 and L4, and the multiresistance gene cfr, were investigated. Furthermore, an epidemiological survey of the nine linezolid-resistant E. faecalis isolates was performed by pulsed field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and DiversiLab. The three methods were compared to evaluate their merits and demerits, respectively. We failed to find the resistance mechanisms that have been revealed in recent years by PCR and sequencing analysis in the linezolid-resistant E. faecalis. Epidemiological investigation suggested that a small-scale outbreak of linezolid-resistant E. faecalis emerged in neurosurgery ICU from March to May of 2013. DiversiLab was a reliable typing tool and a suitable alternative to PFGE because it was as discriminatory as PFGE and better than MLST.


Asunto(s)
Antibacterianos/farmacología , Farmacorresistencia Bacteriana , Enterococcus faecalis/efectos de los fármacos , Infecciones por Bacterias Grampositivas/epidemiología , Infecciones por Bacterias Grampositivas/microbiología , Linezolid/farmacología , China/epidemiología , Infección Hospitalaria , Farmacorresistencia Bacteriana/genética , Electroforesis en Gel de Campo Pulsado , Enterococcus faecalis/clasificación , Enterococcus faecalis/genética , Enterococcus faecalis/aislamiento & purificación , Genes Bacterianos , Humanos , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Mutación
17.
Clin Microbiol Infect ; 19(10): E473-82, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23718188

RESUMEN

A single strain of Mycobacterium massiliense (BRA 100), a subspecies of the Mycobacterium abscessus complex, has been responsible for an epidemic of post-surgical infections in Brazil. Outside Brazil, this is the first report to describe a single emerging strain of M. massiliense (TPE 101) associated with extrapulmonary infections. This phenomenon may be underestimated because sophisticated molecular typing of M. abscessus is not routinely performed. Our molecular epidemiology study was triggered by an outbreak investigation. Nine case isolates were grown from the surgical sites of nine mostly paediatric patients receiving operations from 2010 to 2011. All available non-duplicated isolates of M. abscessus during this period were obtained for comparison. Mycobacteria were characterized by multilocus sequence analysis (MLSA), repetitive sequence PCR (rep-PCR) and pulsed-field gel electrophoresis (PFGE). Of 58 isolates of M. abscessus overall, 56 were clinical isolates. MLSA identified 36 of the isolates as M. massiliense. All case isolates were indistinguishable by PFGE and named the TPE 101 pulsotype. Of the stored strains of M. abscessus, TPE 101 strains were over-represented among the control surgical wound (7/7, 100%) and subcutaneous tissue isolates (4/5, 80%) but rare among the respiratory isolates (1/16, 6%) and absent from external skin, ocular and environmental samples. In conclusion, a unique strain of M. massiliense has emerged as a distinctive pathogen causing soft tissue infections in Taiwan. Further study to identify whether this is due to an occult common source or to specific virulence factors dictating tissue tropism is warranted.


Asunto(s)
Infecciones por Mycobacterium/microbiología , Micobacterias no Tuberculosas/aislamiento & purificación , Infección de la Herida Quirúrgica/microbiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Bacteriemia/epidemiología , Bacteriemia/microbiología , Brasil/epidemiología , Niño , Preescolar , Brotes de Enfermedades/estadística & datos numéricos , Femenino , Humanos , Lactante , Masculino , Persona de Mediana Edad , Epidemiología Molecular , Tipificación Molecular , Infecciones por Mycobacterium/epidemiología , Micobacterias no Tuberculosas/clasificación , Micobacterias no Tuberculosas/genética , Complicaciones Posoperatorias/epidemiología , Complicaciones Posoperatorias/microbiología , Estudios Retrospectivos , Infección de la Herida Quirúrgica/epidemiología , Taiwán/epidemiología
18.
Diagn Microbiol Infect Dis ; 77(4): 337-40, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-24074766

RESUMEN

The typing of multidrug-resistant Acinetobacter baumannii isolates is important for the control and prevention of hospital outbreaks. This study aimed to analyze the molecular epidemiology of 46 OXA-23 carbapenemase-producing A. baumannii strains and compare them to previously described local and international clones (ICs). Isolates were recovered during May 2009-August 2011, from 8 different hospitals in the state of Parana (Brazil). The molecular profiles were determined by repetitive extragenic palindromic PCR. Seven different clusters were identified (A to G). Thirty-two isolates were clustered in the same pattern (clone A), which belong to IC 4.


Asunto(s)
Infecciones por Acinetobacter/epidemiología , Acinetobacter baumannii/clasificación , Acinetobacter baumannii/genética , Infección Hospitalaria , beta-Lactamasas/genética , Infecciones por Acinetobacter/microbiología , Acinetobacter baumannii/aislamiento & purificación , Brasil/epidemiología , ADN Bacteriano , Farmacorresistencia Bacteriana Múltiple/genética , Hospitales , Humanos , Filogenia , Reacción en Cadena de la Polimerasa/métodos , Secuencias Repetitivas de Ácidos Nucleicos , beta-Lactamasas/biosíntesis
19.
Clin Microbiol Infect ; 19(8): 737-42, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23034071

RESUMEN

To investigate the species distribution within the Acinetobacter calcoaceticus-Acinetobacter baumannii complex and the molecular epidemiology of A. baumannii and Acinetobacter nosocomialis, 376 Acinetobacter isolates were collected prospectively from hospitalized patients at 15 medical centres in Germany during three surveillance studies conducted over a 5-year period. Species identification was performed by molecular methods. Imipenem minimum inhibitory concentrations (MIC) were determined by broth microdilution. The prevalence of the most common carbapenemase-encoding genes was investigated by oxacillinase (OXA) -multiplex polymerase chain reaction (PCR). The molecular epidemiology was investigated by repetitive sequence-based PCR (rep-PCR; DiversiLab™). Acinetobacter pittii was the most prevalent Acinetobacter species (n = 193), followed by A. baumannii (n = 140), A. calcoaceticus (n = 10) and A. nosocomialis (n = 8). The majority of A. baumannii was represented by sporadic isolates (n = 70, 50%) that showed unique rep-PCR patterns, 25 isolates (18%) clustered with one or two other isolates, and only 45 isolates (32%) belonged to one of the previously described international clonal lineages. The most prevalent clonal lineage was international clone (IC) 2 (n = 34) and IC 1 (n = 6). According to CLSI, 25 A. baumannii isolates were non-susceptible to imipenem (MIC ≥ 8 mg/L), all of which produced an OXA-58-like or OXA-23-like carbapenemase. The rate of imipenem susceptibility among A. baumannii isolates decreased from 96% in 2005 to 76% in 2009. All other Acinetobacter isolates were susceptible to imipenem. The population structure of carbapenem-susceptible A. baumannii in Germany is highly diverse. Imipenem non-susceptibility was strongly associated with the clonal lineages IC 2 and IC 1. These data underscore the high clonality of carbapenem-resistant A. baumannii isolates.


Asunto(s)
Infecciones por Acinetobacter/epidemiología , Infecciones por Acinetobacter/microbiología , Acinetobacter/clasificación , Acinetobacter/genética , Acinetobacter/aislamiento & purificación , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Alemania/epidemiología , Hospitales , Humanos , Imipenem/farmacología , Pruebas de Sensibilidad Microbiana , Epidemiología Molecular , Tipificación Molecular , Reacción en Cadena de la Polimerasa , Estudios Prospectivos , beta-Lactamasas/genética
20.
Artículo en Inglés | WPRIM | ID: wpr-48263

RESUMEN

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) are important pathogens causing nosocomial infections in Korean hospitals. This study aimed to investigate the epidemiological and genetic diversity of clinical S. aureus isolates in healthcare settings from 2001 to 2008. METHODS: Samples and data were obtained from 986 individuals as part of the National Antimicrobial Surveillance Project, involving 10 regions nationwide. Molecular typing studies, including multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing were performed, and a representative clone of Korean MRSA was classified by combinational grouping using a DiversiLab (DL; bioMérieux, France) repetitive element polymerase chain reaction (rep-PCR) system. RESULTS: Nine Korean MRSA clones (KMRSA-1 to -9) were identified by analysis of genetic backgrounds and molecular characteristics. KMRSA-1 to -3, expressing clonal complex (CC) 5 (carrying SCCmec II), CC8 (carrying SCCmec III), and CC72 (carrying SCCmec IV) were spread nationwide. In contrast, KMRSA-6 was highly prevalent in Gyeongsangnam-do, and KMRSA-4 was highly prevalent in Jeollanam-do and Jeollabuk-do. CONCLUSIONS: Epidemic KMRSA clones were genetically similar to major clones identified from the USA, with the exception of KMRSA-2, which had the SCCmec III type. Our results provide important insights into the distribution and molecular genetics of MRSA strains in Korea and may aid in the monitoring of MRSA spread throughout the country.


Asunto(s)
Humanos , Proteínas Bacterianas/genética , ADN Bacteriano/genética , Electroforesis en Gel de Campo Pulsado , Hospitales , Staphylococcus aureus Resistente a Meticilina/genética , Tipificación de Secuencias Multilocus , Reacción en Cadena de la Polimerasa Multiplex , Prevalencia , República de Corea/epidemiología , Infecciones Estafilocócicas/diagnóstico
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