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@inproceedings{778559,
author = {Kahánková, Jana and Rittich, Bohuslav and Španová, Alena and Petráš, Petr and Růžičková, Vladislava and Doškař, Jiří and Pantůček, Roman},
address = {Cairns, Australia},
booktitle = {13th International Symposium on Staphylococci and Staphylococcal Infections},
keywords = {Staphylococcus aureus; bacteriophages; molecular diagnostics; PCR},
language = {eng},
location = {Cairns, Australia},
pages = {83-83},
publisher = {Australian Society for Antimicrobials},
title = {Molecular diagnostics of temperate bacteriophages of Staphylococcus aureus},
url = {http://www.isssi2008.com/},
year = {2008}
}
TY - JOUR
ID - 778559
AU - Kahánková, Jana - Rittich, Bohuslav - Španová, Alena - Petráš, Petr - Růžičková, Vladislava - Doškař, Jiří - Pantůček, Roman
PY - 2008
TI - Molecular diagnostics of temperate bacteriophages of Staphylococcus aureus
PB - Australian Society for Antimicrobials
CY - Cairns, Australia
KW - Staphylococcus aureus
KW - bacteriophages
KW - molecular diagnostics
KW - PCR
UR - http://www.isssi2008.com/
N2 - Objective: Pathogenic Staphylococcus strains differ in the presence of virulence factors that are encoded mainly by mobile genetic elements, in particular by prophages integrated in the bacterial chromosomes. The study objective was to develop a method for rapid and simple characterization of S. aureus prophages. Methods: The prophages were induced from lysogenic strains by UV-irradiation. They were picked from one plaque and propagated on a non-lysogenic strain to obtain a low titre phage lysate (10e3 PFU/ml). A new method for phage DNA extraction from small volumes (150 microL) 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, antirepressors, amidases and virulence associated genes for Panton-Valentine leukocidin, exfoliative toxin A and those of innate immune evasion cluster. Results: Under optimized induction conditions, prophages were induced from 50 S. aureus strains. The PCR-ready DNA was isolated using new methods and amplified by PCR using newly designed primer sets. The results enabled us to divide the phages into several groups (numbers in brackets) according to capsid structure (9), integrases dictating the attachment site on the host chromosome (10), antirepressor (9), and lytic module (4). We propose updating the phage nomenclature to correspond better to the genomic loci and extensive mosaic pattern of phage genomes. Conclusion: 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. The characterization of staphylococcal prophages is essential for understanding the role of phages in virulence and evolution of S. aureus strains.
ER -
KAHÁNKOVÁ, Jana, Bohuslav RITTICH, Alena ŠPANOVÁ, Petr PETRÁŠ, Vladislava RŮŽIČKOVÁ, Jiří DOŠKAŘ and Roman PANTŮČEK. Molecular diagnostics of temperate bacteriophages of $<$I$>$Staphylococcus aureus$<$/I$>$. In \textit{13th International Symposium on Staphylococci and Staphylococcal Infections}. Cairns, Australia: Australian Society for Antimicrobials, 2008, p.~83.
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