Preparation of a speciesspecific molecular probe for
identification of Staphylococcus aureus

J 2000

Preparation of a speciesspecific molecular probe for identification of Staphylococcus aureus

ŠTĚPÁN, Jiří; Roman PANTŮČEK; Vladislava RŮŽIČKOVÁ; Stanislav ROSYPAL; Jiří DOŠKAŘ et al.

Základní údaje

Originální název

Preparation of a speciesspecific molecular probe for identification of Staphylococcus aureus

Autoři

ŠTĚPÁN, Jiří ; Roman PANTŮČEK ; Vladislava RŮŽIČKOVÁ; Stanislav ROSYPAL a Jiří DOŠKAŘ

Vydání

Scripta medica, Brno, Lékařská fakulta Masarykovy univerzity, 2000, 1211-3395

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

Genetika a molekulární biologie

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/00:00002298

Organizační jednotka

Přírodovědecká fakulta

Změněno: 2. 1. 2001 16:28, prof. RNDr. Roman Pantůček, Ph.D.

Anotace

V originále

In our previous work a 44-kb genomic SmaI-restriction fragment common to all S. aureus strains has been described. This restriction fragment was used as a starting point in preparation of a species-specific hybridisation probe for rapid molecular identification S. aureus strains. EcoRI-restricted genomic DNAs isolated from 13 different S. aureus strains, including the type strain S. aureus CCM 885, and from the type strain S. epidermidis CCM 2124 (negative control) were hybridised to a probe prepared from the whole sequence of the 44-kb fragment. The EcoRI-restriction fragments showing strong hybridisation to the S. aureus 44-kb probe were cloned in the pUCl8 vector in E. coli and used as candidates for the preparation of molecular probes specific for S. aureus. The cloned 2-kb EcoRI-restriction fragment was chosen as the most specific fragment that hybridised to the homologic EcoRl-fragment present in genomic DNA of more than 100 S. aureus strains of different provenance. No hybridisation was detected on genomic DNA isolated from S. epidennidis and the other species of the genus Sraphylococcus. The 2-kb EcoRl-fragment was sequenced and the primers for rapid PCR-identification of S. aureus strains were derived. In using these primers, 825 b PCR amplification products were obtained in DNA isolated from 100 S. aureus strains. No PCR amplification products were obtained in DNA isolated from the other Staphylococcus species.

Návaznosti

GA301/99/D075, projekt VaV

Název: Genotypová diagnostika a typizace klinicky významných koaguláza negativních stafylokoků

Investor: Grantová agentura ČR, Genotypová diagnostika a typizace klinicky významných koaguláza negativních stafylokoků

MSM 143100008, záměr

Název: Genomy a jejich funkce

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Genomy a jejich funkce

Citovat

ŠTĚPÁN, Jiří; Roman PANTŮČEK; Vladislava RŮŽIČKOVÁ; Stanislav ROSYPAL a Jiří DOŠKAŘ. Preparation of a speciesspecific molecular probe for identification of Staphylococcus aureus. Scripta medica. Brno: Lékařská fakulta Masarykovy univerzity, 2000, roč. 72, č. 8, s. 380. ISSN 1211-3395.

@article{327193,
   author = {Štěpán, Jiří and Pantůček, Roman and Růžičková, Vladislava and Rosypal, Stanislav and Doškař, Jiří},
   article_location = {Brno},
   article_number = {8},
   language = {eng},
   issn = {1211-3395},
   journal = {Scripta medica},
   title = {Preparation of a speciesspecific molecular probe for identification of Staphylococcus aureus},
   volume = {72},
   year = {2000}
}

TY  - JOUR
ID  - 327193
AU  - Štěpán, Jiří - Pantůček, Roman - Růžičková, Vladislava - Rosypal, Stanislav - Doškař, Jiří
PY  - 2000
TI  - Preparation of a speciesspecific molecular probe for identification of Staphylococcus aureus
JF  - Scripta medica
VL  - 72
IS  - 8
SP  - 380
EP  - 380
PB  - Lékařská fakulta Masarykovy univerzity
SN  - 12113395
N2  - In our previous work a 44-kb genomic SmaI-restriction fragment common to all S. aureus strains has been described. This restriction fragment was used as a starting point in preparation of a species-specific hybridisation probe for rapid molecular identification S. aureus strains. EcoRI-restricted genomic DNAs isolated from 13 different S. aureus strains, including the type strain S. aureus CCM 885, and from the type strain S. epidermidis CCM 2124 (negative control) were hybridised to a probe prepared from the whole sequence of the 44-kb fragment. The EcoRI-restriction fragments showing strong hybridisation to the S. aureus 44-kb probe were cloned in the pUCl8 vector in E. coli and used as candidates for the preparation of molecular probes specific for S. aureus. The cloned 2-kb EcoRI-restriction fragment was chosen as the most specific fragment that hybridised to the homologic EcoRl-fragment present in genomic DNA of more than 100 S. aureus strains of different provenance. No hybridisation was detected on genomic DNA isolated from S. epidennidis and the other species of the genus Sraphylococcus. The 2-kb EcoRl-fragment was sequenced and the primers for rapid PCR-identification of S. aureus strains were derived. In using these primers, 825 b PCR amplification products were obtained in DNA isolated from 100 S. aureus strains. No PCR amplification products were obtained in DNA isolated from the other Staphylococcus species.
ER  -

ŠTĚPÁN, Jiří; Roman PANTŮČEK; Vladislava RŮŽIČKOVÁ; Stanislav ROSYPAL a Jiří DOŠKAŘ. Preparation of a speciesspecific molecular probe for identification of Staphylococcus aureus. \textit{Scripta medica}. Brno: Lékařská fakulta Masarykovy univerzity, 2000, roč.~72, č.~8, s.~380. ISSN~1211-3395.

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