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Genotype variability of TSST-1 positive <I>Staphylococcus aureus</I> strains in the Czech Republic

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HRSTKA Roman RŮŽIČKOVÁ Vladislava PANTŮČEK Roman PETRÁŠ Petr DOŠKAŘ Jiří

Rok publikování 2001
Druh Článek ve sborníku
Konference V. Pracovní setkání biochemiků a molekulárních biologů
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
Obor Genetika a molekulární biologie
Popis Species Staphylococcus aureus belongs to clinically important gram-positive cocci. Some S. aureus strains produce TSST-1 toxin, which causes human disease called Toxic Shock Syndrom (TSS). Two forms of TSS were differentiated: the first type occurs in connection with menses, the second one is formed of cases, when TSS is a complication of another staphylococcal disease, for exampte food poisoning. Data about abiGty of S. aureus strains to produce TSST-1 toxin are clearly different. Therefore, we analyzed genomic DNAs of 26 S. aureus strains isolated from patients in the Czech Republic in connection with TSS. Staphylococcal isolates were analyzed by PCR using specific oligonucleotide primers to confirm presence of specific sequences of tst gene. To determine genetic variability, we performed macrorestriction analysis using PFGE, which enables determination of 20 different PFGE types. Strains were divided into 8 PFGE groups (ET1-ET8) according to Smal restriction patteros based on the similarity level between 65 - l00%. Majority of strains were classified into the group ET7, in whicb 14 strains were included (67% from all analyzed TSST-1 positive strains). The next method was PCR-amplification of variable-length 16S-23S rDNA spacer regions. The analyzed strains were distributed into 4 groups (SP1-SP4). Ztie distribution of strains into groups SP1-SP4 confirmed results gained by RFLP. In the next part of our work, we identified localization of tss specific sequences. The tst sequence was locatized on staphylococcal chromosome using hybridization by means of specific probe to SmaI DNA restriction patterns of S. aureus strains. Positive signals conesponded with ability of the strains to grow in Tip free medium, what indicates linkage of Trp operon and tst gene.
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