KAHÁNKOVÁ, Jana, Roman PANTŮČEK, Vladislava RŮŽIČKOVÁ and Jiří DOŠKAŘ. Multiplex PCR strategy for characterization of modular genomic structure of Staphylococcus aureus bacteriophages. In Pathophysiology of Staphylococci. Abstract book. Würzburg, Germany: Transregional Collaborative Research Center 34. p. 63. 2008.
Other formats:   BibTeX LaTeX RIS
Basic information
Original name Multiplex PCR strategy for characterization of modular genomic structure of Staphylococcus aureus bacteriophages
Name in Czech Technika multiplex PCR pro charakterizaci modulární struktury genomu bakteriofágů Staphylococcus aureus
Authors KAHÁNKOVÁ, Jana (203 Czech Republic), Roman PANTŮČEK (203 Czech Republic, guarantor), Vladislava RŮŽIČKOVÁ (203 Czech Republic) and Jiří DOŠKAŘ (203 Czech Republic).
Edition Abstract book. Würzburg, Germany, Pathophysiology of Staphylococci, p. 63-63, 2008.
Publisher Transregional Collaborative Research Center 34
Other information
Original language English
Type of outcome Proceedings paper
Field of Study Genetics and molecular biology
Country of publisher Germany
Confidentiality degree is not subject to a state or trade secret
WWW URL
RIV identification code RIV/00216224:14310/08:00026648
Organization unit Faculty of Science
Keywords in English Staphylococcus aureus; bacteriophages; molecular diagnostics; mobile genetic elements
Tags bacteriophages, mobile genetic elements, Molecular diagnostics, Staphylococcus aureus
Tags International impact
Changed by Changed by: prof. RNDr. Roman Pantůček, Ph.D., učo 842. Changed: 31/10/2008 14:52.
Abstract
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.
Abstract (in Czech)
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.
Links
MSM0021622415, plan (intention)Name: Molekulární podstata buněčných a tkáňových regulací
Investor: Ministry of Education, Youth and Sports of the CR, Molecular basis of cell and tissue regulations
PrintDisplayed: 19/4/2024 11:45