Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 19 de 19
Filtrar
Mais filtros










Base de dados
Tipo de estudo
Intervalo de ano de publicação
1.
Int J Mol Sci ; 21(12)2020 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-32545831

RESUMO

Trueperella pyogenes is an important opportunistic animal pathogen. Different antimicrobials, including aminoglycosides, are used to treat T. pyogenes infections. The aim of the present study was to evaluate aminoglycoside susceptibility and to detect aminoglycoside resistance determinants in 86 T. pyogenes isolates of different origin. Minimum inhibitory concentration of gentamicin, streptomycin, and kanamycin was determined using a standard broth microdilution method. Genetic elements associated with aminoglycoside resistance were investigated by PCR and DNA sequencing. All studied isolates were susceptible to gentamicin, but 32.6% and 11.6% of them were classified as resistant to streptomycin and kanamycin, respectively. A total of 30 (34.9%) isolates contained class 1 integrons. Class 1 integron gene cassettes carrying aminoglycoside resistance genes, aadA11 and aadA9, were found in seven and two isolates, respectively. Additionally, the aadA9 gene found in six isolates was not associated with mobile genetic elements. Moreover, other, not carried by gene cassettes, aminoglycoside resistance genes, strA-strB and aph(3')-IIIa, were also detected. Most importantly, this is the first description of all reported genes in T. pyogenes. Nevertheless, the relevance of the resistance phenotype to genotype was not perfectly matched in 14 isolates. Therefore, further investigations are needed to fully explain aminoglycoside resistance mechanisms in T. pyogenes.

2.
Foods ; 8(7)2019 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-31331094

RESUMO

The prevalence of Bacillus cereus in a total of 585 samples of food products (herbs and spices, breakfast cereals, pasta, rice, infant formulas, pasteurized milk, fresh acid and acid/rennet cheeses, mold cheeses and ripening rennet cheeses) marketed in Poland was investigated. The potential of 1022 selected isolates of B. cereus to hydrolyze casein, starch and tributyrin, to ferment lactose, to grow at 7 C/10 days, to produce Nhe and Hbl toxin and to possess the ces gene was verified. B. cereus was found in 38.8% of the analyzed samples, reaching levels from 0.3 to 3.8 log CFU g-1 or mL-1. From the 1022 isolates, 48.8%, 36.0%, 98.9%, 80.0% and 25.0% were capable of fermenting lactose, producing amylase, protease, lipase and growing at 7 C/10 days, respectively, indicating spoilage potentiality. The occurrence of toxigenic B. cereus strains in all tested market products, both of plant (55.8% Hbl(+), 70.7% Nhe(+) and 1.7% ces(+) isolates) and animal origin (84.9% Hbl(+), 82.7% Nhe(+) and 0.9% ces(+) isolates) indicates the possible risk of foodborne infections/intoxications that occur as a result of the possibility of the development of B. cereus in favorable conditions and consumption of these products.

3.
Int J Mol Sci ; 20(11)2019 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-31167367

RESUMO

Bacteria from the species Trueperella pyogenes are a part of the biota of skin and mucous membranes of the upper respiratory, gastrointestinal, or urogenital tracts of animals, but also, opportunistic pathogens. T. pyogenes causes a variety of purulent infections, such as metritis, mastitis, pneumonia, and abscesses, which, in livestock breeding, generate significant economic losses. Although this species has been known for a long time, many questions concerning the mechanisms of infection pathogenesis, as well as reservoirs and routes of transmission of bacteria, remain poorly understood. Pyolysin is a major known virulence factor of T. pyogenes that belongs to the family of cholesterol-dependent cytolysins. Its cytolytic activity is associated with transmembrane pore formation. Other putative virulence factors, including neuraminidases, extracellular matrix-binding proteins, fimbriae, and biofilm formation ability, contribute to the adhesion and colonization of the host tissues. However, data about the pathogen-host interactions that may be involved in the development of T. pyogenes infection are still limited. The aim of this review is to present the current knowledge about the pathogenic potential and virulence of T. pyogenes.


Assuntos
Actinomycetaceae/fisiologia , Infecções por Bactérias Gram-Positivas/microbiologia , Actinomycetaceae/classificação , Actinomycetaceae/patogenicidade , Animais , Toxinas Bacterianas/química , Toxinas Bacterianas/genética , Toxinas Bacterianas/imunologia , Reservatórios de Doenças , Genoma Bacteriano , Genômica/métodos , Infecções por Bactérias Gram-Positivas/imunologia , Infecções por Bactérias Gram-Positivas/transmissão , Interações Hospedeiro-Patógeno/imunologia , Humanos , Filogenia , Ligação Proteica , Domínios e Motivos de Interação entre Proteínas , RNA Ribossômico 16S , Relação Estrutura-Atividade , Virulência , Fatores de Virulência/química , Fatores de Virulência/genética , Fatores de Virulência/imunologia
4.
J Vet Res ; 62(1): 57-64, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29978128

RESUMO

Introduction: One aim of the study was to evaluate the impact when added to feed of the two potentially probiotic strains of lactic acid bacteria (LAB) Lactobacillus plantarum K KKP 593/p and Lactobacillus rhamnosus KKP 825 on production performance, health, and the composition of gut microbiota. The complementary aim was to assess the safety of these strains in broiler rearing. Material and Methods: A total of 500 one-day-old Ross 308 chicks were divided into four groups. The experimental factor was the admixture of bacterial preparation to the feed at different doses: the recommended maximum dose, a dose ten times higher, the recommended minimum dose, and a zero dose for the control group not receiving bacteria. Results: Addition of bacteria to the diets did not have a significant effect on the final body weight, final body weight gain, nor total feed intake or feed conversion. However, lactic acid bacteria had a positive effect on chicken health. Mortality among chickens fed with LAB was reduced. Moreover, LAB feeding inhibited the growth of Salmonella spp. and Clostridium perfringens in the intestines. There were no significant differences in chicken performance by dose of bacteria in the feed. The group dosed with LAB ten times higher than the recommended maximum did not demonstrate changes in biochemical or haematological parameters of blood compared to the remaining groups. Conclusion: Feeding chicken broilers with two potentially probiotic LAB strains is safe and impacts animal health positively.

5.
Vet Microbiol ; 216: 25-30, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29519521

RESUMO

A total of 43 Campylobacter isolates from poultry, cattle and pigs were investigated for their ability to form biofilm. The studied strains were also screened for motility, adhesion and invasion of Caco-2 cells as well as extracellular DNase activity. The relation between biofilm formation and selected phenotypes was examined. Biofilm formation by the tested strains was found as irrespective from their motility and not associated with colonization abilities of human Caco-2 cells. Results of our study show that Campylobacter isolates from various animal sources are able to form biofilm and invade human Caco-2 cells in vitro.


Assuntos
Infecções por Campylobacter/veterinária , Campylobacter/genética , Campylobacter/isolamento & purificação , Fenótipo , Animais , Aderência Bacteriana , Biofilmes/crescimento & desenvolvimento , Células CACO-2 , Campylobacter/classificação , Campylobacter/patogenicidade , Infecções por Campylobacter/microbiologia , Bovinos , Galinhas , Humanos , Reação em Cadeia da Polimerase , Aves Domésticas/microbiologia , Doenças das Aves Domésticas/microbiologia , Suínos , Fatores de Virulência
6.
Food Microbiol ; 65: 221-230, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28400006

RESUMO

Bacteria of the genus Cronobacter are emerging food-borne pathogens. Foods contaminated with Cronobacter spp. may pose a risk to infants or adults with suppressed immunity. This study was aimed at determining the microbiological quality of ready-to-eat (RTE) plant-origin food products available on the Polish market with special emphasis on the prevalence of Cronobacter genus bacteria. Analyses were carried out on 60 samples of commercial RTE type plant-origin food products, including: leaf vegetables (20 samples), sprouts (20 samples) and non-pasteurized vegetable, fruit and fruit-vegetable juices (20 samples). All samples were determined for the total count of aerobic mesophilic bacteria (TAMB) and for the presence of Cronobacter spp. The isolates of Cronobacter spp. were subjected to genetic identification and differentiation by 16S rDNA sequencing, PCR-RFLP analysis and RAPD-PCR and evaluation of antibiotic susceptibility by the disk diffusion assay. The TAMB count in samples of lettuces, sprouts and non-pasteurized fruit, vegetable and fruit-vegetable juices was in the range of 5.6-7.6, 6.7-8.4 and 2.9-7.7 log CFU g-1, respectively. The presence of Cronobacter spp. was detected in 21 (35%) samples of the products, including in 6 (30%) samples of leaf vegetables (rucola, lamb's lettuce, endive escarola and leaf vegetables mix) and in 15 (75%) samples of sprouts (alfalfa, broccoli, small radish, lentil, sunflower, leek and sprout mix). No presence of Cronobacter spp. was detected in the analyzed samples of non-pasteurized fruit, vegetable and fruit-vegetable juices. The 21 strains of Cronobacter spp. isolated from leaf vegetable and sprouts included: 13 strains of C. sakazakii, 4 strains of C. muytjensii, 2 strains of C. turicensis, one strain of C. malonaticus and one strain of C. condimenti. All isolated C. sakazakii, C. muytjensii, C. turicensis and C. malonaticus strains were sensitive to ampicillin, cefepime, chloramphenicol, gentamycin, streptomycin, tetracycline, ciprofloxacin and cotrimoxazol, whereas the C. condimenti isolate showed intermediate resistance to streptomycin and cotrimoxazole.


Assuntos
Cronobacter/isolamento & purificação , Fast Foods/microbiologia , Sucos de Frutas e Vegetais/microbiologia , Frutas/microbiologia , Verduras/microbiologia , Antibacterianos/farmacologia , Contagem de Colônia Microbiana , Cronobacter/efeitos dos fármacos , Cronobacter/genética , Cronobacter/crescimento & desenvolvimento , Cronobacter sakazakii/efeitos dos fármacos , Cronobacter sakazakii/genética , Cronobacter sakazakii/isolamento & purificação , Farmacorresistência Bacteriana Múltipla , Microbiologia de Alimentos , Qualidade dos Alimentos , Humanos , Medicago sativa/microbiologia , Testes de Sensibilidade Microbiana , Folhas de Planta/microbiologia , Polônia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , RNA Ribossômico 16S/genética , Plântula/microbiologia , Análise de Sequência de DNA
7.
J Vet Med Sci ; 78(4): 543-9, 2016 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-26655770

RESUMO

Rhodococcus equi is the causative agent of rhodococcosis in horses, resulting in significant morbidity and mortality in foals. This bacterium has also been isolated from a variety of animals and is being increasingly reported as a cause of infection in humans, mainly in immunosuppressed individuals. Laboratory diagnostics of R. equi infections based only on conventional microbiological methods shows low accuracy and can lead to misidentification. The objective of the study was to develop and evaluate a real-time PCR assay for direct detection of R. equi in various clinical specimens, including tissue samples. The species-specific region of the gene encoding R. equi cholesterol oxidase, choE, was used as a qPCR-target. The diagnostic applicability of the assay was confirmed by testing various tissue specimens obtained from horses with clinical signs of rhodoccocal infection and swine submaxillary lymph nodes. The rate of R. equi detection in clinical specimens by the developed assay was higher in comparison to the culture method (90% vs. 60.0% of positive samples) and conventional PCR (90.0% vs. 20.0% of positive samples). In case of 13 samples that were negative in the culture-based method, R. equi was detected by the developed assay. Only in one case, it gave negative result for culture-positive sample. The assay may provide a simple and rapid tool to complement the classical methods of R. equi detection based on culture and phenotypic identification of isolates, as the performed evaluation indicated a high specificity and accuracy of the results.


Assuntos
Infecções por Actinomycetales/veterinária , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Rhodococcus equi/isolamento & purificação , Infecções por Actinomycetales/diagnóstico , Infecções por Actinomycetales/microbiologia , Animais , Colesterol Oxidase/análise , Colesterol Oxidase/genética , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/microbiologia , Cavalos , Suínos , Doenças dos Suínos/diagnóstico , Doenças dos Suínos/microbiologia
8.
J Sci Food Agric ; 96(11): 3897-905, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26693837

RESUMO

BACKGROUND: Wheat flour is one of the most common causative agents of food allergy. The study presents the selection and characterization of lactic acid bacteria (LAB) strains capable of hydrolyzing/modifying allergenic proteins of wheat flour. Hydrolysis of wheat proteins was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting with sera from patients with food allergy to gluten. RESULTS: The analysis of electrophoretic profiles of protein extracted from sourdough shows the capability of selected LAB strains for proteolytic degradation of wheat proteins that belong to two factions: albumin/globulin (hydrolysis of 13 polypeptides with a molecular weight between 103 and 22 kDa); and gliadin (seven polypeptides with a molecular weight between 39 and 24 kDa). All analyzed strains were capable of hydrolyzing some IgE-binding epitopes of wheat allergens. The lack of such changes in control samples indicates that they were induced rather by the proteolytic activity of bacterial strains than endogenous enzymes of wheat flour. The gluten proteins were susceptible to hydrolysis by sequential digestion with pepsin and trypsin. CONCLUSION: The selected strains exhibit proteolytic activity, which leads to a reduction in allergenicity of wheat sourdoughs. These strains may be applied as specific starter cultures to prepare bakery products of special nutritional use. © 2015 Society of Chemical Industry.


Assuntos
Antígenos de Plantas/metabolismo , Pão/análise , Dieta Livre de Glúten , Farinha/análise , Glutens/metabolismo , Lactobacillales/metabolismo , Proteínas de Vegetais Comestíveis/metabolismo , Albuminas/efeitos adversos , Albuminas/química , Albuminas/metabolismo , Antígenos de Plantas/efeitos adversos , Antígenos de Plantas/química , Pão/efeitos adversos , Pão/microbiologia , Epitopos/efeitos adversos , Epitopos/metabolismo , Fermentação , Farinha/efeitos adversos , Farinha/microbiologia , Gliadina/efeitos adversos , Gliadina/química , Gliadina/metabolismo , Globulinas/efeitos adversos , Globulinas/química , Globulinas/metabolismo , Glutens/efeitos adversos , Humanos , Lactobacillales/crescimento & desenvolvimento , Lactobacillales/isolamento & purificação , Peso Molecular , Proteínas de Vegetais Comestíveis/efeitos adversos , Proteínas de Vegetais Comestíveis/química , Polônia , Proteólise , Sementes/efeitos adversos , Sementes/química , Sementes/microbiologia , Especificidade da Espécie , Triticum/efeitos adversos , Triticum/química , Triticum/microbiologia , Hipersensibilidade a Trigo/dietoterapia , Hipersensibilidade a Trigo/imunologia
9.
Pol J Microbiol ; 64(3): 285-8, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26638537

RESUMO

Escherichia coli is a common cause of infections in companion animals. In recent years the increasing prevalence of resistance to ß-lactams, including extended-spectrum cephalosporins, antimicrobials frequently used in small animal veterinary practice, was observed in canine isolates of E. coli. The aim of this study was to detect and to characterize extended-spectrum ß-lactamases (ESBLs) produced by E. coli isolated from diseased dogs in Poland. Four isolates out of 119 studied (3.4%) were ESBL-positive. They harbored the bla(SHV-12), bla(CTX-M-15), and bla(TEM-116) genes. This study provides the first report of the occurrence of ESBL-producing E. coli in dogs in Poland.


Assuntos
Doenças do Cão/microbiologia , Infecções por Escherichia coli/veterinária , Proteínas de Escherichia coli/metabolismo , Escherichia coli/enzimologia , beta-Lactamases/metabolismo , Animais , Antibacterianos/farmacologia , Doenças do Cão/epidemiologia , Cães , Farmacorresistência Bacteriana , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/genética , Polônia/epidemiologia , beta-Lactamases/genética
10.
Vet Microbiol ; 172(1-2): 272-8, 2014 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-24878324

RESUMO

Rhodococcus equi is a soil saprophyte and an opportunistic pathogen causing infections in animals, and rarely in humans. The presence of R. equi in tissues and faeces of some wild animal species was demonstrated previously. In this study we characterized R. equi isolates from submaxillary lymph nodes of free-living wild boars (n=23), red deer (n=2) and roe deer (n=2). This is the first description of R. equi strains isolated from tissues of the Cervidae. All isolates were initially recognized as R. equi based on the phenotypic properties. Their identification was confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, detection of the choE gene and by sequence analysis of the 16S rRNA and rpoB genes. The presence of three plasmidic genes (traA, vapA and vapB) associated with R. equi virulence was investigated by PCR. In 16 wild boar isolates the traA and vapB genes were detected and they were located on virulence plasmids type 5, 7 or 11. The isolates from cervids and the remaining wild boar isolates were classified as avirulent based on a genotype traA(-)/vapA(-)B(-). In summary, these results confirm that wild boars can be a source of intermediately virulent R. equi strains, and indicate that red deer and roe deer can be a reservoir of avirulent R. equi strains.


Assuntos
Infecções por Actinomycetales/veterinária , Cervos/microbiologia , Genes Bacterianos , RNA Ribossômico 16S/genética , Rhodococcus equi/genética , Sus scrofa/microbiologia , Infecções por Actinomycetales/epidemiologia , Infecções por Actinomycetales/microbiologia , Infecções por Actinomycetales/transmissão , Animais , Reservatórios de Doenças , Humanos , Linfonodos/microbiologia , Filogenia , Polônia/epidemiologia , Rhodococcus equi/classificação , Rhodococcus equi/isolamento & purificação , Rhodococcus equi/patogenicidade , Suínos , Virulência
11.
Pol J Microbiol ; 62(1): 51-8, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23829077

RESUMO

The aim of this study was to analyze data collected by the SENTINEL influenza surveillance system in Poland in the first post-pandemic season 2010/2011. The results include weeks 35/2010-17/2011. Physicians registered weekly number of influenza-like illnesses and collected specimens. Laboratory tests were done using PCR and/or real-time PCR or immunofluorescence. Laboratories also isolated the influenza virus. Influenza-like illness incidence amounted to 2802.7/100000. Weekly incidence ranged from 11.3/100000 to 232/100000. The most affected group was children 0-4 years. Influenza-like illness peak occurred between weeks 02/2011 and 11/2011. Influenza infections were confirmed in 34.9% of specimens. More cases were caused by influenza A, including A(H1N1)pdm09 than influenza B (59.9% vs. 40.1%). The isolated strains were similar to A/California/7/2009 or B/Brisbane/60/2008. Season 2010/2011 in Poland did not differ from the rest of Europe. Further improvement is necessary, especially in the area of specimen collection at the beginning of an epidemic season and carrying out the isolation of the influenza virus.


Assuntos
Influenza Humana/epidemiologia , Epidemias , Humanos , Vírus da Influenza A Subtipo H1N1 , Influenza Humana/virologia , Polônia/epidemiologia , Estações do Ano , Vigilância de Evento Sentinela , Fatores de Tempo
12.
Arch Virol ; 158(8): 1743-53, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23515874

RESUMO

This study was performed to develop real-time PCR (qPCR) for detection of human seasonal and avian influenza viruses in duplex format. First duplex qPCR detects haemagglutinin (HA) gene of influenza virus A(H1N1)pdm09 and HA gene of influenza virus A(H3N2), the second reaction detects neuraminidase (NA) gene of influenza virus A(H3N2) and NA gene of influenza virus A(H1N1)pdm09 and A(H5N1), and the third reaction detects HA gene of influenza A(H5N1) and nonstructural protein gene of influenza B virus. Primers and probes were designed using multiple alignments of target gene sequences of different reference strains. Assays were optimised for identical thermocycling conditions. Their specificity was confirmed by conventional PCR and monoplex qPCR with nucleic acids isolated from different influenza viruses and other respiratory pathogens. Plasmid constructs with a fragment of specific gene were used to assess sensitivity of the assay. The limit of detection ranged from 27 to 96 cDNA copies/reaction. Clinical specimens (n = 107) have been tested using new assays, immunofluorescence and monoplex qRT-PCR. It has been shown that developed assays have been capable of rapid and accurate simultaneous detection and differentiation of influenza viruses. They are more sensitive than immunofluorescence and at least as sensitive as monoplex qRT-PCR.


Assuntos
Vírus da Influenza A/classificação , Vírus da Influenza A/isolamento & purificação , Influenza Humana/diagnóstico , Influenza Humana/virologia , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Virologia/métodos , Primers do DNA/genética , Humanos , Vírus da Influenza A/genética , Técnicas de Diagnóstico Molecular/métodos , Sondas de Oligonucleotídeos/genética , Sensibilidade e Especificidade
13.
Adv Exp Med Biol ; 755: 237-41, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-22826072

RESUMO

Children are an important vector for spreading influenza and they are at increased risk for complications. The appropriate diagnosis of influenza may help start early antiviral treatment and may optimize the use of antibiotics and additional laboratory tests. The objective of this study was to describe the influence of rapid influenza detection test (RIDT) on clinical management of children with acute febrile respiratory tract infections. The method consisted of a prospective, open, cohort study conducted in three primary care clinics in Warsaw, Poland, during the epidemic influenza seasons of 2009/2010 and 2010/2011. A total number of 256 children of the age 0-5 years with symptoms of febrile respiratory tract infection were enrolled into the study. A 115 of them were tested with RIDT (BD Directigen EZ FluA + B) and another 141 children, who were not tested, constituted a control group. We found that RIDT gave positive results in 35 (30%) out of the 115 tested children. Antibiotics, additional blood tests and urinalysis were administered more often in the control group compared with the rapid test group (16% vs. 7%; 14% vs. 5%, and 47% vs. 32%, respectively). Chest radiograms were made only in six cases of children from the control group. We conclude that in children with symptoms of acute febrile respiratory tract infection, the rapid influenza detection test provides a rational use of antivirals, reduces an inappropriate use of antibiotics, and decreases a number of additional tests conducted.


Assuntos
Febre/tratamento farmacológico , Influenza Humana/diagnóstico , Infecções Respiratórias/tratamento farmacológico , Antibacterianos/uso terapêutico , Antivirais/uso terapêutico , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Influenza Humana/tratamento farmacológico , Masculino
14.
Med Dosw Mikrobiol ; 64(2): 129-37, 2012.
Artigo em Polonês | MEDLINE | ID: mdl-23072058

RESUMO

INTRODUCTION: Among different laboratory techniques used in influenza surveillance, methods based on molecular biology, such as real-time PCR, play increasingly important role. They allow detection and identification of viruses currently circulating in human population and responsible for infections and diseases. The aim of the study was to develop duplex real-time PCR assay for detection and differentiation of influenza virus subtype A(H1N1) pdm09 and A(H3N2). METHODS: Specific primers targeting a highly conservative region ofhaemagglutinin gene and a dual-color TaqMan probes, labeled with fluorophore JOE (560 nm) and quencher BHQ-1 or CalFluor Red 610 (610 nm) and BHQ-2 were designed. The specificity of duplex reaction was evaluated using RNA isolated from reference strains of influenza virus A(H1N1)pdm09, A(H3N2), A(H1N1) and A(H5N1), B and other respiratory pathogens. Sensitivity of the assay was tested using serial dilutions ofplasmid constructs carrying fragment of specific viral HA gene, in range between 10 and 10(5) copies/sample. A number of 116 clinical samples were tested using developed duplex qPCR assay in comparison with the CDC monoplex assay, RESULTS: Positive duplex qPCR results were obtained only in samples containing RNA of influenza viruses of a specific subtype. The limit of detection (LOD) of developed method amounted up to 27 copies/sample for A(H1N1)pdm09 and up to 37 copies/sample for A(H3N2). CONCLUSIONS: The results show that developed TaqMan-based duplex qPCR assay is a valuable diagnostic tool in influenza virus surveillance for using in direct study of clinical specimens as well as identification of isolated strains of influenza virus.


Assuntos
Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Vírus da Influenza A Subtipo H3N2/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sequência de Bases , Vírus da Influenza A Subtipo H1N1/classificação , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H3N2/classificação , Vírus da Influenza A Subtipo H3N2/genética , Dados de Sequência Molecular , Plasmídeos/genética , Especificidade da Espécie
15.
Postepy Hig Med Dosw (Online) ; 66: 452-60, 2012 Jun 28.
Artigo em Polonês | MEDLINE | ID: mdl-22922145

RESUMO

Respiratory viruses contribute to significant morbidity and mortality in healthy and immunocompromised individuals and are considered as a significant economic burden in the healthcare system. The similar clinical symptoms in the course of different viral and bacterial respiratory infections make the proper diagnosis difficult. An accurate and prompt diagnostics is crucial for infection control and patient management decisions, especially regarding the use of antibacterial or antiviral therapy and hospitalization. Moreover, the identification of the causative agent eliminates inappropriate use of antibiotics and may reduce the cost of healthcare. A wide variety of diagnostic procedures is applied for the detection of viral agents responsible for respiratory tract infections. For many years, the viral antigen detection and standard isolation technique in cell culture was the main method used in routine diagnostics. However, in recent years the nucleic acid amplification techniques have become widely used and have significantly improved the sensitivity of viral detection in clinical specimens. Molecular diagnostic assays have contributed to revealing high rates of co-infection (multiplex reactions) and allow identification of agents that are difficult to culture. This paper discusses a number of technical aspects of the current most commonly used techniques, their general principles, main benefits and diagnostic value, but also some of their limitations.


Assuntos
Infecções Respiratórias/diagnóstico , Infecções Respiratórias/virologia , Vírus/isolamento & purificação , Antígenos Virais/isolamento & purificação , Humanos , Técnicas de Amplificação de Ácido Nucleico/métodos , Vírus/genética , Vírus/imunologia
16.
Vet Microbiol ; 160(1-2): 69-76, 2012 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-22658663

RESUMO

Trueperella (Arcanobacterium) pyogenes is an opportunistic animal pathogen, which in European bison is associated with different suppurative infections mainly of the urogenital tract. Little is known about the virulence of this bacterium and about the pathogenesis of infections. The main objective of this study was to determine phenotypic properties and virulence genotypes of the twenty-five T. pyogenes strains isolated from lesions in various tissues of free-living European bison. Classical bacteriological methods were used for phenotypic characterization. Genes encoding seven known and putative virulence factors of T. pyogenes were detected by PCR technique. Analysis of 16S rDNA partial sequences was performed to establish phylogenetic relationships of the isolated strains. All isolates showed typical morphological features of T. pyogenes and variable biochemical activity. Most of them displayed a strong positive effect in synergistic CAMP test. For all isolates the 16S rRNA gene partial sequence was identical to that of the T. pyogenes reference strain. All isolates carried the plo and fimA genes, while the nanH, nanP, cbpA, fimC and fimG genes were present in 40, 44, 12, 88 and 24% of the isolates, respectively. The T. pyogenes strains isolated from European bison represented various phenotypes and virulence genotypes, but there was no association between the investigated properties of the bacteria and the type of anatomopathological lesions from which they were isolated. These results indicate that the studied virulence factors of T. pyogenes are not significant determinants of the localization and type of infection caused by this bacterium.


Assuntos
Infecções por Actinomycetales/veterinária , Arcanobacterium/genética , Arcanobacterium/patogenicidade , Bison , Infecções por Actinomycetales/microbiologia , Animais , Arcanobacterium/isolamento & purificação , DNA Ribossômico/genética , Feminino , Genótipo , Masculino , Filogenia , Reação em Cadeia da Polimerase , Virulência/genética , Fatores de Virulência/genética
17.
Przegl Epidemiol ; 65(2): 199-203, 2011.
Artigo em Polonês | MEDLINE | ID: mdl-21913459

RESUMO

A total number of 1,081,974 cases of influenza and influenza-like illness were registered in Poland in 2009 (incidence 2,835.9 per 100,000 population). It was nearly 5 times more than in 2008. The impact on increase of the number of reported cases have had two factors: the pandemic of influenza caused by virus A(H1N1)v, and increasing of the surveillance sensitivity. 3,177 (0.29%) cases was laboratory confirmed. In the area of particular regions incidence ranged from 805.2 in swietokrzyskie voivodeship to 5,257.9 in warmifisko-mazurskie voivodeship. Nearly 37% of cases were children under 15 years. The incidence in this age group was 6,851.2 (from 2,010.1 in Swietokrzyskie voivodeship, to 13,291.6 in warminsko-mazurskie voivodeship). The highest reported incidence was observed in age group 5-14 years (7,135.2). To hospitals, mainly for epidemiological reasons, 8,944 people were sent (0.83% all cases). According to Central Statistical Office data, there were 87 death cases, including 8 (9.2%) children in the age of 15. 70.1% of deaths were registered as cases caused by identified influenza virus.


Assuntos
Surtos de Doenças/estatística & dados numéricos , Influenza Humana/diagnóstico , Influenza Humana/epidemiologia , Sistema de Registros/estatística & dados numéricos , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Incidência , Lactente , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Vírus da Influenza A Subtipo H3N2/isolamento & purificação , Vacinas contra Influenza/administração & dosagem , Influenza Humana/prevenção & controle , Influenza Humana/virologia , Influenzavirus A/isolamento & purificação , Masculino , Vacinação em Massa/estatística & dados numéricos , Pessoa de Meia-Idade , Polônia/epidemiologia , População Rural/estatística & dados numéricos , População Urbana/estatística & dados numéricos , Adulto Jovem
18.
J Clin Microbiol ; 49(6): 2216-21, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21471335

RESUMO

Mixed infections of a single host with different variants of influenza A virus are the main source of reassortants which may have unpredictable properties when they establish themselves in the human population. In this report we describe a method for rapid detection of mixed influenza virus infections with the seasonal A/H1N1 human strain and the pandemic A/H1N1/v strain which emerged in 2009 in Mexico and the United States. The influenza virus A/H1N1 variants were characterized by the multitemperature single-stranded conformational polymorphism (MSSCP) method. The MSSCP gel patterns of hemagglutinin gene fragments of pandemic A/H1N1/v and different seasonal A/H1N1 strains were easily distinguishable 2 h after completion of reverse transcription-PCR (RT-PCR). Using the MSSCP-based genotyping approach, coinfections with seasonal and pandemic variants of the A/H1N1 subtype were identified in 4 out of 23 primary samples obtained from patients that presented with influenza-like symptoms to hospitals across Poland during the 2009-2010 epidemic season. Pandemic influenza virus strain presence was confirmed in all these primary samples by real-time RT-PCR. The sensitivity level of the MSSCP-based minor genetic variant detection was 0.1%, as determined on a mixture of DNA fragments obtained from amplification of the hemagglutinin gene of seasonal and pandemic strains. The high sensitivity of the method suggests its applicability for characterization of new viral variants long before they become dominant.


Assuntos
Vírus da Influenza A Subtipo H1N1/classificação , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Influenza Humana/diagnóstico , Influenza Humana/virologia , Polimorfismo Conformacional de Fita Simples , Virologia/métodos , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Genótipo , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Humanos , Vírus da Influenza A Subtipo H1N1/genética , Masculino , Pessoa de Meia-Idade , Polônia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sensibilidade e Especificidade , Adulto Jovem
19.
Pol J Microbiol ; 57(2): 105-12, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18646397

RESUMO

Phenotypic approaches based on metabolic and biological characteristics of Corynebacterium pseudotuberculosis have been limited due to insufficient discrimination between closely related isolates. In this paper we present performance and convenience of three molecular typing methods: BOX-PCR, random amplification of polymorphic DNA (RAPD) and amplification of DNA fragments surrounding rare restriction site (ADSRRS-fingerprinting) in genome analysis of these bacteria. Among examined 61 strains there were distinguished four, eight and 10 different genotypes by BOX-PCR, RAPD and ADSRRS-fingerprinting, respectively. The value of discrimination index was the lowest for BOX-PCR (D = 0.265), much bigger for RAPD (D = 0.539) and the highest for ADSRRS-fingerprinting (D = 0.604). The good discriminatory ability and reproducibility of RAPD and ADSRRS-fingerprinting indicates that those techniques may be particularly applied for epidemiological studies of C. pseudotuberculosis isolates. We found that ADSRRS-fingerprinting is a rapid method offering good discrimination power, excellent reproducibility and may be applied for epidemiological studies of intraspecific genetic relatedness of C. pseudotuberculosis strains.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/classificação , Impressões Digitais de DNA/métodos , Doenças das Cabras/microbiologia , Linfadenite/veterinária , Animais , Infecções por Corynebacterium/microbiologia , Corynebacterium pseudotuberculosis/genética , Primers do DNA , DNA Bacteriano/genética , Genótipo , Cabras , Linfadenite/microbiologia , Polônia , Reação em Cadeia da Polimerase , Técnica de Amplificação ao Acaso de DNA Polimórfico , Especificidade da Espécie
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA