Nejvíce citovaný článek - PubMed ID 16681147
Multiplex PCR for detection of three exfoliative toxin serotype genes in Staphylococcus aureus
The aim of this study was to describe the prevalence of different Staphylococcus species isolated from pathological processes and lesions in domestic animals in the Czech Republic and to detect and describe oxacillin (methicillin)-resistant strains (MRS). During the years 2019-2020, a total of 5218 veterinary clinical samples from the Czech Republic were tested. Testing was performed by culture methods and typing by molecular phenotypic methods MALDI-TOF MS and PCR. Antimicrobial susceptibility testing of the strains was performed by the disk diffusion method. A total of 854 staphylococci strains were identified (16.37% prevalence), out of which 43 strains of 6 species of staphylococci were MRS (n = 43; 0.82% prevalence). Of the MRS strains, the most prevalent species were Staphylococcus pseudintermedius (n = 24; 0.46% prevalence) and Staphylococcus aureus (n = 7; 0.13% prevalence). Susceptibility testing showed resistance to beta-lactam antibiotics and, depending on the species, also to trimethoprim/sulfamethoxazole, gentamicin, tetracycline, erythromycin, clindamycin, and enrofloxacin. For further characterization of MRS, PCR assay for virulence factor genes was performed. Seven of the 14 target genes were observed only in S. aureus, except for the eno gene encoding laminin-binding protein, which was also detected in other staphylococci. It is necessary to emphasize the issue of correct using of antimicrobials in practice and antibiotic policy in university teaching and to create stricter legislation that would prevent the widespread use of antimicrobials in veterinary medicine, especially in livestock to reduce the emergence and spread of antimicrobial resistance.
- Klíčová slova
- occurrence, organs, prevalence, susceptibility, veterinary medicine, virulence genes,
- Publikační typ
- časopisecké články MeSH
Exfoliative toxin A (ETA)-coding temperate bacteriophages are leading contributors to the toxic phenotype of impetigo strains of Staphylococcus aureus. Two distinct eta gene-positive bacteriophages isolated from S. aureus strains which recently caused massive outbreaks of pemphigus neonatorum in Czech maternity hospitals were characterized. The phages, designated ϕB166 and ϕB236, were able to transfer the eta gene into a prophageless S. aureus strain which afterwards converted into an ETA producer. Complete phage genome sequences were determined, and a comparative analysis of five designed genomic regions revealed major variances between them. They differed in the genome size, number of open reading frames, genome architecture, and virion protein patterns. Their high mutual sequence similarity was detected only in the terminal regions of the genome. When compared with the so far described eta phage genomes, noticeable differences were found. Thus, both phages represent two new lineages of as yet not characterized bacteriophages of the Siphoviridae family having impact on pathogenicity of impetigo strains of S. aureus.
- MeSH
- DNA virů chemie genetika MeSH
- DNA viry genetika izolace a purifikace MeSH
- epidemický výskyt choroby MeSH
- exfoliatiny genetika MeSH
- fylogeneze MeSH
- genom virový * MeSH
- impetigo epidemiologie mikrobiologie MeSH
- infekce spojené se zdravotní péčí epidemiologie MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- novorozenec MeSH
- otevřené čtecí rámce MeSH
- polymorfismus délky restrikčních fragmentů MeSH
- pořadí genů MeSH
- porodnice MeSH
- přenos genů horizontální MeSH
- profágy klasifikace genetika izolace a purifikace MeSH
- sekvenční analýza DNA MeSH
- sekvenční homologie MeSH
- shluková analýza MeSH
- stafylokokové bakteriofágy klasifikace genetika izolace a purifikace MeSH
- stafylokokové infekce epidemiologie mikrobiologie MeSH
- Staphylococcus aureus izolace a purifikace virologie MeSH
- syntenie MeSH
- transdukce genetická MeSH
- Check Tag
- lidé MeSH
- novorozenec MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika epidemiologie MeSH
- Názvy látek
- DNA virů MeSH
- exfoliatiny MeSH
A multiplex PCR assay was developed for the detection of toxigenic and pathogenic V. cholerae from direct water sources using specific primers targeting diverse genes, viz. outer membrane protein (ompW), cholera toxin (ctxB), ORF specific for O1 (rfbG), zonula occludens (zot) and toxin co-regulated pilus (tcpB); among these genes, ompW acts as internal control for V. cholerae, the ctx gene as a marker for toxigenicity and tcp for pathogenicity. The sensitivity of multiplex PCR was 5 x 10(4) V. cholerae cells per reaction. The procedure was simplified as direct bacterial cells were used as template and there was no need for DNA extraction. The assay was specific as no amplification occurred with the other bacteria used. Toxigenic V. cholerae were artificially spiked in different water samples, filtered through a 0.45 microm membrane, and the filters containing bacteria were enriched in APW for 6 h. PCR following filtration and enrichment could detect as little as 8 V. cholerae cells per mL in different spiked water samples. Various environmental potable water samples were screened for the presence of V. cholerae using this assay procedure. The proposed method is rapid, sensitive and specific for environmental surveillance for the presence of toxigenic-pathogenic and nonpathogenic V. cholerae.
- MeSH
- bakteriální proteiny genetika MeSH
- cholerový toxin genetika MeSH
- DNA primery MeSH
- látky znečišťující vodu analýza MeSH
- monitorování životního prostředí metody MeSH
- polymerázová řetězová reakce metody MeSH
- senzitivita a specificita MeSH
- sladká voda mikrobiologie MeSH
- Vibrio cholerae non-O1 * genetika izolace a purifikace patogenita MeSH
- Vibrio cholerae O1 * genetika izolace a purifikace patogenita MeSH
- zásobování vodou analýza MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- Názvy látek
- bakteriální proteiny MeSH
- cholerový toxin MeSH
- DNA primery MeSH
- látky znečišťující vodu MeSH
The occurrence of Staphylococcus aureus in rabbit feces, cecum and meat and its enterotoxin production, susceptibility to antibiotics and its sensitivity or resistance to bacteriocins produced by enterococci with probiotic properties were determined. Isolates were resistant to ampicillin, penicillin, phosphomycin and methicillin; a high percentage of susceptibility was also recorded to vancomycin, chloramphenicol, tetracycline and tobramycin. S. aureus isolates did not produce enterotoxins and were sensitive to partially purified enterocins (PPB) EK13, AL41 and EF2019 in the range of 100 to 12800 AU/mL; all S. aureus isolates, except the strain SA 2A/3, exhibited the highest sensitivity to PPB EK13. On the other hand, all strains were resistant to PPB CCM4231.
- MeSH
- bakteriociny metabolismus MeSH
- cékum mikrobiologie MeSH
- feces mikrobiologie MeSH
- hospodářská zvířata mikrobiologie MeSH
- králíci MeSH
- maso mikrobiologie MeSH
- mikrobiální testy citlivosti MeSH
- mnohočetná bakteriální léková rezistence * MeSH
- přemostěné cyklické sloučeniny metabolismus MeSH
- rezistence na methicilin * MeSH
- Staphylococcus aureus účinky léků izolace a purifikace MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Slovenská republika MeSH
- Názvy látek
- bakteriociny MeSH
- enterocin MeSH Prohlížeč
- přemostěné cyklické sloučeniny MeSH
A simple and specific method for direct detection of bovine mastitis pathogens (Streptococcus agalactiae (GBS), Staphylococcus aureus and Escherichia coli) in milk products, bacterial samples from milk and isolated bacterial DNA was developed. The method is based on polymerase chain reaction (PCR) using sequence-specific primers only for GBS and species-specific primers derived from 16S and 23S rRNA for all chosen species. The presence of the gene of surface immunogenic protein (Sip) in bovine GBS isolates, described previously only in human GBS isolates was confirmed. The GBS detection was performed with the sequence coding for surface immunogenic protein from GBS human isolates designated as Sip specific sequence (SSS); this sequence was selected for specific primer design. The sequence is unique for GBS and was designed from a consensus of all known sip genes. The specific identification was shown on a collection of 75 GBS bovine isolates from different localities in Slovakia. All isolates were positive to SSS, 16S and 23S rRNA sequence. The 16S and 23S rRNA PCR detection was also performed with S. aureus and E. coli isolates and specific PCR products were also detected. The detection limit of this assay for milk products was 6 CFU/microL (i.e. 6000 CFU/mL) for GBS and E. coli, and 16 CFU/microL for S. aureus. This rapid, sensitive and specific diagnostic method can be performed within hours and represents an innovative diagnostic tool for the detection of milk pathogens in dairy products.
- MeSH
- antigeny bakteriální genetika MeSH
- bakteriologické techniky * MeSH
- DNA bakterií analýza genetika MeSH
- DNA primery MeSH
- elektroforéza v agarovém gelu MeSH
- Escherichia coli genetika izolace a purifikace MeSH
- mléčné výrobky mikrobiologie MeSH
- molekulární sekvence - údaje MeSH
- polymerázová řetězová reakce metody MeSH
- potravinářská mikrobiologie MeSH
- ribozomální DNA analýza genetika MeSH
- RNA ribozomální 16S genetika MeSH
- RNA ribozomální 23S genetika MeSH
- sekvence nukleotidů MeSH
- senzitivita a specificita MeSH
- Staphylococcus aureus genetika izolace a purifikace MeSH
- Streptococcus agalactiae genetika izolace a purifikace MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antigeny bakteriální MeSH
- DNA bakterií MeSH
- DNA primery MeSH
- ribozomální DNA MeSH
- RNA ribozomální 16S MeSH
- RNA ribozomální 23S MeSH
- SIP protein, Streptococcus group B MeSH Prohlížeč
We describe bacteriophage therapy in the case of a healthcare worker whose gastrointestinal tract was colonized by methicillin-resistant Staphylococcus aureus (MRSA) with subsequent urinary tract infection caused by the same pathogen. Oral treatment with anti-MRSA phages resulted in eradication of the carrier status.
- MeSH
- bakteriofágy * MeSH
- dospělí MeSH
- lidé MeSH
- přenašečství prevence a kontrola MeSH
- přenos infekce ze zdravotnického pracovníka na pacienta prevence a kontrola MeSH
- rezistence na methicilin * MeSH
- stafylokokové infekce prevence a kontrola terapie virologie MeSH
- Staphylococcus aureus účinky léků virologie MeSH
- zdravotní sestry MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- práce podpořená grantem MeSH