PCR fingerprinting
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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 212 type and reference strains deposited in the Czech Collection of Microorganisms (Brno, Czech Republic) and covering 41 Staphylococcus species comprising 21 subspecies was characterised using rep-PCR fingerprinting with the (GTG)₅ primer in order to evaluate this method for identification of staphylococci. All strains were typeable using the (GTG)₅ primer and generated PCR products ranging from 200 to 4500 bp. Numerical analysis of the obtained fingerprints revealed (sub)species-specific clustering corresponding with the taxonomic position of analysed strains. Taxonomic position of selected strains representing the (sub)species that were distributed over multiple rep-PCR clusters was verified and confirmed by the partial rpoB gene sequencing. Staphylococcus caprae, Staphylococcus equorum, Staphylococcus sciuri, Staphylococcus piscifermentans, Staphylococcus xylosus, and Staphylococcus saprophyticus revealed heterogeneous fingerprints and each (sub)species was distributed over several clusters. However, representatives of the remaining Staphylococcus spp. were clearly separated in single (sub)species-specific clusters. These results showed rep-PCR with the (GTG)₅ primer as a fast and reliable method applicable for differentiation and straightforward identification of majority of Staphylococcus spp.
- MeSH
- DNA bakterií genetika MeSH
- DNA fingerprinting metody MeSH
- DNA primery genetika MeSH
- DNA řízené RNA-polymerasy chemie MeSH
- fylogeneze MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- podjednotky proteinů genetika MeSH
- polymerázová řetězová reakce metody MeSH
- shluková analýza MeSH
- stafylokokové infekce mikrobiologie MeSH
- Staphylococcus klasifikace genetika izolace a purifikace MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
A group of lactobacilli isolated from the cervix of 31 healthy women was characterized by (GTG)(5)-polymerase chain reaction (PCR) fingerprinting in order to evaluate this method for identification of vaginal lactobacilli. Obtained fingerprints were compared with profiles available in an in-house database of the CCM bacteria collection covering type and reference strains of multiple lactic acid bacteria including lactobacilli. Selected strains representing individual clusters were further identified by pheS gene sequencing. In total, six lactobacillus species were found among lactobacilli isolated from the cervix of healthy women. The (GTG)(5)-PCR method identified Lactobacillus gasseri (11 strains), Lactobacillus fermentum (one), and some of the Lactobacillus jensenii strains (eight out of 11), but failed to identify the remaining strains, including the Lactobacillus crispatus (18), Lactobacillus mucosae (one), and Lactobacillus vaginalis (one) species. L. jensenii strains were distributed over two fingerprint clusters. The majority of samples was dominated by one (GTG)(5)-PCR type. The rep-PCR fingerprinting using the (GTG)(5) primer allowed straightforward identification of many, but not all, isolates. This method has been shown to be a useful tool for fast screening and grouping of vaginal lactobacilli, but its combination with another identification method is needed to obtain reliable identification results. In addition, Lactobacillus acidophilus was not shown to be the most common inhabitant of the female genital tract as generally assumed.
- MeSH
- cervix uteri mikrobiologie MeSH
- DNA bakterií genetika MeSH
- DNA fingerprinting metody MeSH
- DNA primery genetika MeSH
- dospělí MeSH
- fylogeneze MeSH
- Lactobacillus klasifikace genetika izolace a purifikace MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- polymerázová řetězová reakce metody MeSH
- techniky typizace bakterií MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
Analysis of actA gene sequence polymorphism has been shown to be an effective and relatively inexpensive method for subtyping Listeria monocytogenes isolates, allowing the division of the population of this species into two deeply separate lineages. This sequence-based method as well as PCR-mediated fingerprinting were applied here for the differentiation of 49 isolates of food and clinical origin. Correlation between these two typing approaches was high. Both methods divided the isolates into two lineages, designated I (33 isolates) and II (16 isolates). All the 33 lineage I isolates were assigned to the same, or closely related, six clusters by both typing methods. For the lineage II isolates, PCR fingerprinting was found to be more discriminatory. The isolates were characterized by cell invasion assay. All highly invasive isolates were assigned to lineage I, which constituted a heterogeneous group also containing low-invasive isolates. High-invasive isolates were not found in the genetically determined lineage II. A particular actA cluster, designated Ha, contained all the isolates showing the lowest invasiveness. A common trait of the isolates belonging to this cluster was the presence of a threonine-441 of the deduced ActA sequence instead of the alanine-441 present in the remaining isolates. Thirteen human isolates were classified to lineage I and five to lineage II. A PCR-based method can therefore differentiate L. monocytogenes isolates in accordance with the current phylogenetic model of the evolution of this species.
- MeSH
- bakteriální proteiny genetika MeSH
- cytologické techniky MeSH
- DNA fingerprinting metody MeSH
- fylogeneze MeSH
- lidé MeSH
- Listeria monocytogenes genetika izolace a purifikace klasifikace MeSH
- listeriové infekce mikrobiologie MeSH
- membránové proteiny genetika MeSH
- nádorové buněčné linie MeSH
- polymerázová řetězová reakce MeSH
- potravinářská mikrobiologie MeSH
- techniky typizace bakterií metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
Gene expression profiles are important data to reveal the functions of genes putatively involved in crucial biological processes. RNA arbitrarily primed polymerase chain reaction (RAP-PCR) and specifically primed reverse transcription polymerase chain reaction (RT-PCR) were combined to screen differentially expressed genes following development of a commercial Bacillus thuringiensis subsp. kurstaki strain 8010 (serotype 3a3b). Six differentially expressed transcripts (RAP1 to RAP6) were obtained. RAP1 encoded a putative triple helix repeat-containing collagen or an exosporium protein H related to spore pathogenicity. RAP2 was homologous to a ClpX protease and an ATP-dependent protease La (LonB), which likely acted as virulence factors. RAP3 was homologous to a beta subunit of propionyl-CoA carboxylase required for the development of Myxococcus xanthus. RAP4 had homology to a quinone oxidoreductase involved in electron transport and ATP formation. RAP5 showed significant homology to a uridine kinase that mediates phosphorylation of uridine and azauridine. RAP6 shared high sequence identity with 3-methyl-2-oxobutanoate-hydroxymethyltransferase (also known as ketopantoate hydroxymethyltransferase or PanB) involved in the operation of the tricarboxylic acid cycle. The findings described here would help to elucidate the molecular mechanisms underlying the differentiation process of B. thuringiensis and unravel novel pathogenic genes.
- MeSH
- Bacillus thuringiensis klasifikace genetika růst a vývoj izolace a purifikace MeSH
- bakteriální proteiny genetika metabolismus MeSH
- DNA fingerprinting metody MeSH
- polymerázová řetězová reakce s reverzní transkripcí metody MeSH
- regulace genové exprese u bakterií MeSH
- spory bakteriální klasifikace genetika růst a vývoj metabolismus MeSH
- vývojová regulace genové exprese MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
