Detection and quantification of Staphylococcus aureus genes and
mobile genetic elements within bacteriophage particles by qPCR

a 2012

Detection and quantification of Staphylococcus aureus genes and mobile genetic elements within bacteriophage particles by qPCR

MAŠLAŇOVÁ, Ivana, Jiří DOŠKAŘ, Marian VARGA, Lucie KUNTOVÁ, Jan MUŽÍK et. al.

Základní údaje

Originální název

Detection and quantification of Staphylococcus aureus genes and mobile genetic elements within bacteriophage particles by qPCR

Autoři

MAŠLAŇOVÁ, Ivana, Jiří DOŠKAŘ, Marian VARGA, Lucie KUNTOVÁ, Jan MUŽÍK, Denisa MALÚŠKOVÁ, Vladislava RŮŽIČKOVÁ a Roman PANTŮČEK

Vydání

15 th International Symposium on Staphylococci and Staphylococcal Infections (ISSSI2012), Lyon, France, 2012

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

Genetika a molekulární biologie

Stát vydavatele

Francie

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

Staphylococcus aureus; bacteriophage; horizontal gene transfer; qPCR

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 22. 11. 2012 13:11, prof. RNDr. Roman Pantůček, Ph.D.

Anotace

V originále

Rapid evolution of Staphylococcus species contributes to substantial variability of strains and formation of novel clones and is facilitated by the acquisition of new genes through horizontal gene transfer (HGT). The newly acquired genes responsible for virulence and ATB resistance are rapidly disseminated in the staphylococcal population. It is widely accepted that temperate bacteriophages play a major role in this process and contribute to spread of mobile genetic elements (MGE) including SCCmec element. Methicillin resistant S. aureus strains (MRSA) represent a significant medical problem. In the present day, 11 different types of SCCmec element are described, which carry mecA gene responsible for methicillin resistance, but the transfer mechanisms and evolution of SCCmec element are still discussed. In this study, we developed and optimized the technique for detection and quantification of S. aureus bacterial genes directly within the phage particles via qPCR. The packaging frequencies of 12 different bacterial genes including genes localized on staphylococcal cassette chromosome mec (SCCmec type I), staphylococcal pathogenicity island SaPI1 and genomic islands were compared. Non-parametric statistical analysis showed that transducing staphylococcal bacteriophages 11, 80 and 80alpha of serogroup B, in contrast to serogroup A bacteriophage 81, efficiently package selected chromosomal genes localized in 4 various loci of the chromosome and genes carried on MGE like SCCmec type 1, SaPI1, genomic islands Saalpha and Sabeta, and plasmids with various frequency. Bacterial gene copy number per ng (CG/ng) of DNA isolated from phage particles was established for all 12 analysed genes. The new and crucial finding that serogroup B bacteriophages can package concurrently ccrA1 and mecA localized on SCCmec type I into their capsids indicates that generalized transduction plays an important role in the evolution and emergence of novel MRSA clones.

Návaznosti

ED1.1.00/02.0068, projekt VaV

Název: CEITEC - central european institute of technology

GA310/09/0459, projekt VaV

Název: Úloha bakteriofágů v horizontálním přenosu genů virulence a rezistence u Staphylococcus aureus

Investor: Grantová agentura ČR, Úloha bakteriofágů v horizontálním přenosu genů virulence a rezistence u Staphylococcus aureus

NT12395, projekt VaV

Název: Molekulární průkaz a analýza invazivních kmenů small colony variants (SCV) a rezistentních kmenů S. aureus od pacientů s cystickou fibrózou

Citovat

MAŠLAŇOVÁ, Ivana, Jiří DOŠKAŘ, Marian VARGA, Lucie KUNTOVÁ, Jan MUŽÍK, Denisa MALÚŠKOVÁ, Vladislava RŮŽIČKOVÁ a Roman PANTŮČEK. Detection and quantification of Staphylococcus aureus genes and mobile genetic elements within bacteriophage particles by qPCR. In 15 th International Symposium on Staphylococci and Staphylococcal Infections (ISSSI2012), Lyon, France. 2012.

@proceedings{1074378,
   author = {Mašlaňová, Ivana and Doškař, Jiří and Varga, Marian and Kuntová, Lucie and Mužík, Jan and Malúšková, Denisa and Růžičková, Vladislava and Pantůček, Roman},
   booktitle = {15 th International Symposium on Staphylococci and Staphylococcal Infections (ISSSI2012), Lyon, France},
   keywords = {Staphylococcus aureus; bacteriophage; horizontal gene transfer; qPCR},
   language = {eng},
   title = {Detection and quantification of Staphylococcus aureus genes and mobile genetic elements within bacteriophage particles by qPCR},
   url = {http://isssi2012.univ-lyon1.fr/en},
   year = {2012}
}

TY  - CONF
ID  - 1074378
AU  - Mašlaňová, Ivana - Doškař, Jiří - Varga, Marian - Kuntová, Lucie - Mužík, Jan - Malúšková, Denisa - Růžičková, Vladislava - Pantůček, Roman
PY  - 2012
TI  - Detection and quantification of Staphylococcus aureus genes and mobile genetic elements within bacteriophage particles by qPCR
KW  - Staphylococcus aureus
KW  - bacteriophage
KW  - horizontal gene transfer
KW  - qPCR
UR  - http://isssi2012.univ-lyon1.fr/en
N2  - Rapid evolution of Staphylococcus species contributes to substantial variability of strains and formation of novel clones and is facilitated by the acquisition of new genes through horizontal gene transfer (HGT). The newly acquired genes responsible for virulence and ATB resistance are rapidly disseminated in the staphylococcal population. It is widely accepted that temperate bacteriophages play a major role in this process and contribute to spread of mobile genetic elements (MGE) including SCCmec element. Methicillin resistant S. aureus strains (MRSA) represent a significant medical problem. In the present day, 11 different types of SCCmec element are described, which carry mecA gene responsible for methicillin resistance, but the transfer mechanisms and evolution of SCCmec element are still discussed. In this study, we developed and optimized the technique for detection and quantification of S. aureus bacterial genes directly within the phage particles via qPCR. The packaging frequencies of 12 different bacterial genes including genes localized on staphylococcal cassette chromosome mec (SCCmec type I), staphylococcal pathogenicity island SaPI1 and genomic islands were compared. Non-parametric statistical analysis showed that transducing staphylococcal bacteriophages 11, 80 and 80alpha of serogroup B, in contrast to serogroup A bacteriophage 81, efficiently package selected chromosomal genes localized in 4 various loci of the chromosome and genes carried on MGE like SCCmec type 1, SaPI1, genomic islands Saalpha and Sabeta, and plasmids with various frequency. Bacterial gene copy number per ng (CG/ng) of DNA isolated from phage particles was established for all 12 analysed genes. The new and crucial finding that serogroup B bacteriophages can package concurrently ccrA1 and mecA localized on SCCmec type I into their capsids indicates that generalized transduction plays an important role in the evolution and emergence of novel MRSA clones.
ER  -

MAŠLAŇOVÁ, Ivana, Jiří DOŠKAŘ, Marian VARGA, Lucie KUNTOVÁ, Jan MUŽÍK, Denisa MALÚŠKOVÁ, Vladislava RŮŽIČKOVÁ a Roman PANTŮČEK. Detection and quantification of Staphylococcus aureus genes and mobile genetic elements within bacteriophage particles by qPCR. In \textit{15 th International Symposium on Staphylococci and Staphylococcal Infections (ISSSI2012), Lyon, France}. 2012.

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