2008
Multiplex PCR strategy for characterization of modular genomic structure of Staphylococcus aureus bacteriophages
KAHÁNKOVÁ, Jana, Roman PANTŮČEK, Vladislava RŮŽIČKOVÁ a Jiří DOŠKAŘZákladní údaje
Originální název
Multiplex PCR strategy for characterization of modular genomic structure of Staphylococcus aureus bacteriophages
Název česky
Technika multiplex PCR pro charakterizaci modulární struktury genomu bakteriofágů Staphylococcus aureus
Autoři
KAHÁNKOVÁ, Jana (203 Česká republika), Roman PANTŮČEK (203 Česká republika, garant), Vladislava RŮŽIČKOVÁ (203 Česká republika) a Jiří DOŠKAŘ (203 Česká republika)
Vydání
Abstract book. Würzburg, Germany, Pathophysiology of Staphylococci, s. 63-63, 2008
Nakladatel
Transregional Collaborative Research Center 34
Další údaje
Jazyk
angličtina
Typ výsledku
Stať ve sborníku
Obor
Genetika a molekulární biologie
Stát vydavatele
Německo
Utajení
není předmětem státního či obchodního tajemství
Odkazy
Kód RIV
RIV/00216224:14310/08:00026648
Organizační jednotka
Přírodovědecká fakulta
Klíčová slova anglicky
Staphylococcus aureus; bacteriophages; molecular diagnostics; mobile genetic elements
Příznaky
Mezinárodní význam
Změněno: 31. 10. 2008 14:52, prof. RNDr. Roman Pantůček, Ph.D.
V originále
Pathogenic Staphylococcus aurues strains differ in the presence of virulence factors that are encoded mainly by mobile genetic elements, in particular by prophages. The study objective was to develop a method for rapid and simple characterization of S. aureus prophages. The prophages were induced from lysogenic strains by UV-irradiation. Phages were picked up from one plaque and propagated on a non-lysogenic strain to obtain a low titre phage lysate (10 e3 PFU/ml). A new method for phage DNA extraction from small volumes of low titre phage lysate was developed using magnetic nonporous microspheres P(HEMA-co-EDMA) and NucleoMag. The phage DNAs were characterized by multiplex PCR assays targeting capsid genes (portal and tail), genes for phage integrases, anti-repressors, amidases and virulence associated genes for Panton-Valentine leukocidin, exfoliative toxin A and those of innate immune evasion cluster. The PCR-ready DNA was isolated using novel method and amplified by PCR using newly designed primer sets. The results enabled us to divide the phage genomic modules into several types (numbers in brackets): capsid structure (9), integrases dictating the attachment site on the host chromosome (10), anti-repressor (10), and lytic module (4). We propose updating the phage nomenclature to correspond better to the genomic loci and extensive mosaic pattern of phage genomes. The rapid and simple method for DNA extraction followed by PCR based diagnosis of phage genomic modules is helpful in effective study of phage dynamics.
Česky
Pathogenic Staphylococcus aurues strains differ in the presence of virulence factors that are encoded mainly by mobile genetic elements, in particular by prophages. The study objective was to develop a method for rapid and simple characterization of S. aureus prophages. The prophages were induced from lysogenic strains by UV-irradiation. Phages were picked up from one plaque and propagated on a non-lysogenic strain to obtain a low titre phage lysate (10 e3 PFU/ml). A new method for phage DNA extraction from small volumes of low titre phage lysate was developed using magnetic nonporous microspheres P(HEMA-co-EDMA) and NucleoMag. The phage DNAs were characterized by multiplex PCR assays targeting capsid genes (portal and tail), genes for phage integrases, anti-repressors, amidases and virulence associated genes for Panton-Valentine leukocidin, exfoliative toxin A and those of innate immune evasion cluster. The PCR-ready DNA was isolated using novel method and amplified by PCR using newly designed primer sets. The results enabled us to divide the phage genomic modules into several types (numbers in brackets): capsid structure (9), integrases dictating the attachment site on the host chromosome (10), anti-repressor (10), and lytic module (4). We propose updating the phage nomenclature to correspond better to the genomic loci and extensive mosaic pattern of phage genomes. The rapid and simple method for DNA extraction followed by PCR based diagnosis of phage genomic modules is helpful in effective study of phage dynamics.
Návaznosti
| MSM0021622415, záměr |
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