RAPD
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DNA fingerprinting methods, RAPD with 7 random primers, and rep-PCR using both BOXA1R and (GTG)(5) ones, were used for the discrimination of 16 type and collection Bifidobacterium strains of 9 species of human origin, B. animalis ssp. animalis and B. animalis ssp. lactis and 7 Bifidobacterium strains collected in the Culture Collection of Dairy Microorganisms (CCDM). Both RAPD and rep-PCR methods provided similar results. The strains were identified as B. animalis ssp. lactis (6 strains) and B. adolescentis (1 strain). The reclassification of the collection strain CCM 3761 as B. pseudocatenulatum species (previously classified as B. adolescentis) was confirmed.
- MeSH
- Bifidobacterium genetika izolace a purifikace klasifikace MeSH
- DNA bakterií genetika MeSH
- DNA fingerprinting metody MeSH
- fylogeneze MeSH
- lidé MeSH
- polymerázová řetězová reakce metody MeSH
- technika náhodné amplifikace polymorfní DNA metody MeSH
- techniky typizace bakterií metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
A group of 67 Lactobacillus spp. strains containing Lactobacillus casei/paracasei, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus plantarum, Lactobacillus rhamnosus and Lactobacillus salivarius species isolated from early childhood caries and identified to the species level in a previous study (Svec et al., Folia Microbiol 54:53-58, 2009) was characterized by automated ribotyping performed by the RiboPrinter microbial characterization system and by randomly amplified polymorphic DNA fingerprinting (RAPD-PCR) with M13 primer to evaluate these techniques for characterization of lactobacilli associated with dental caries. Ribotyping revealed 55 riboprints among the analysed group. The automatic identification process performed by the RiboPrinter system identified 18 strains to the species level, however cluster analysis divided obtained ribotype patterns into individual clusters mostly corresponding to the species assignment of particular strains. RAPD-PCR fingerprints revealed by the individual Lactobacillus spp. showed higher variability than the ribotype patterns and the fingerprint profiles generated by the analysed species were distributed among one to four clusters. In conclusion, ribotyping is shown to be more convenient for the identification purposes while RAPD-PCR fingerprinting results indicate this method is a better tool for typing of Lactobacillus spp. strains occurring in dental caries.
- MeSH
- automatizace MeSH
- genotyp MeSH
- Lactobacillus klasifikace izolace a purifikace MeSH
- lidé MeSH
- ribotypizace metody MeSH
- shluková analýza MeSH
- technika náhodné amplifikace polymorfní DNA metody MeSH
- techniky typizace bakterií metody MeSH
- zubní kaz mikrobiologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
A single random oligonucleotide 3H primer has been previously applied in random-amplified- polymorphic-DNA (RAPD)-PCR to distinguish stocked bacteria E. coli within a cocktail mixture also containing Enterococcus faecalis, Bifidobacterium longum and Ruminococcus gnavus. In this study, we demonstrate that a 702 base pair (bp) gene fragment can be amplified as a unique pattern by RAPD-PCR using a 3H primer in human faeces containing E. coli. This unique 702 bp amplicon contained a 687 bp gene fragment identified as the C-terminal region of the glutamate-ammonia-ligase adenyltransferase (glnE) gene of E. coli. By high-resolution melt (HRM) analysis, a mean melt-curve temperature of this 702 bp amplicon was determined to be approximately 88.1 ± 0.22 degrees Celsius (°C). A combination of RAPD with HRM in one single reaction based on this amplicon can achieve semi-quantitative detection of up to 102 CFU/ml of E. coli. To increase the signal intensity of HRM, a primer pair capable of screening E. coli directly from fresh human faeces was re-designed from the 687 bp gene segment, giving a mean peak melt-curve temperature at 88.35 ± 0.11 °C. Finally, single-nucleotide polymorphisms of this 687 bp gene segment were analysed for pathogenic E. coli strains, including UMN026, O83:H1, O104:H4, O157:H7 and O169:H41. We conclude that this 687 bp segment of the glnE gene has a high potential for screening of human faecal E. coli, including pathogenic strains, in contaminated food and water.
- MeSH
- bakteriální geny * MeSH
- DNA primery metabolismus MeSH
- Escherichia coli enzymologie genetika izolace a purifikace MeSH
- feces mikrobiologie MeSH
- glutaminsynthetasa chemie genetika metabolismus MeSH
- jednonukleotidový polymorfismus genetika MeSH
- lidé MeSH
- párování bází genetika MeSH
- sekvence aminokyselin MeSH
- sekvence nukleotidů MeSH
- technika náhodné amplifikace polymorfní DNA * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The aim of this study was to analyse genotypes, antimicrobial susceptibility patterns and serotypes in Pseudomonas aeruginosa clinical strains, including the clonal dissemination of particular strains throughout various intensive care units in one medical centre. Using random amplified polymorphic DNA (RAPD-PCR) and P. aeruginosa antisera, 22 different genotypes and 8 serotypes were defined among 103 isolates from 48 patients. No direct association between P. aeruginosa strain genotypes and serotypes was observed. RAPD typing in strains with the same serotype revealed different genotypes and, on the contrary, most strains with a different serotype displayed the same amplification pattern. The resulting banding patterns showed a high degree of genetic heterogeneity among all isolates from the patients examined, suggesting a non-clonal relationship between isolates from these patients. A higher degree of antibiotic resistance and stronger biofilm production in common genotypes compared to rare ones and genetic homogeneity of the most resistant strains indicated the role of antibiotic pressure in acquiring resistant and more virulent strains in our hospital. In conclusion, genetic characterisation of P. aeruginosa strains using RAPD method was shown to be more accurate in epidemiological analyses than phenotyping.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence MeSH
- biofilmy * MeSH
- dospělí MeSH
- genotyp MeSH
- jednotky intenzivní péče statistika a číselné údaje MeSH
- lidé středního věku MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- mladý dospělý MeSH
- pseudomonádové infekce mikrobiologie MeSH
- Pseudomonas aeruginosa účinky léků genetika izolace a purifikace fyziologie MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- technika náhodné amplifikace polymorfní DNA MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
The genus Vibrio is characterized by a large number of species and some of them are human pathogens causing gastrointestinal and wound infections through the ingestion or manipulation of contaminated fishes and shellfish including Vibrio parahaemolyticus and Vibrio alginolyticus. In this study, we reported the phenotypic and molecular characterization of 9 V. parahaemolyticus and 27 V. alginolyticus strains isolated from outbreaks affecting cultured Gilthead sea bream (Sparus aurata L.) and Sea bass (Dicentrarchus labrax) along the Tunisian coast from 2008 to 2009. All isolates were tested for the presence of DNase, caseinase, protease, lipase, amylase, gelatinase, hemolytic activity and antibacterial resistance to different drugs. Randomly amplified polymorphic DNA was used to examine the genetic relatedness among the V. parahaemolyticus and V. alginolyticus strains.
- MeSH
- DNA fingerprinting MeSH
- epidemický výskyt choroby veterinární MeSH
- fenotyp MeSH
- infekce bakteriemi rodu Vibrio epidemiologie mikrobiologie veterinární MeSH
- mořan zlatý mikrobiologie MeSH
- nemoci ryb epidemiologie mikrobiologie MeSH
- Percoidea mikrobiologie MeSH
- technika náhodné amplifikace polymorfní DNA metody MeSH
- Vibrio alginolyticus genetika izolace a purifikace MeSH
- Vibrio parahaemolyticus genetika izolace a purifikace MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Tunisko MeSH
- MeSH
- Aspergillus izolace a purifikace klasifikace MeSH
- automatizované zpracování dat metody využití MeSH
- DNA primery genetika MeSH
- finanční podpora výzkumu jako téma MeSH
- klinické laboratorní techniky metody využití MeSH
- lidé MeSH
- technika náhodné amplifikace polymorfní DNA metody statistika a číselné údaje využití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- fylogeneze MeSH
- Giardia lamblia genetika izolace a purifikace kontraindikace MeSH
- krysa rodu rattus MeSH
- polymerázová řetězová reakce MeSH
- psi MeSH
- technika náhodné amplifikace polymorfní DNA MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- psi MeSH
- zvířata MeSH