Wykorzystanie techniki PCR-RFLP do wykrywania mutacji w kodonie 86 genu gyrA zwiazanej z opornoscia na fluorochinolony paleczek Campylobacter jejuni

• Autorzy: Wardak S. , Szych J. , Cieslik A.

Warianty tytułu

EN

PCR-restriction fragment length polymorphism assay [PCR-RFLP] as an useful tool for detection of mutation in gyrA gene at 86-Thr position associated with fluoroquinolone resistance in Campylobacter jejuni

• Języki publikacji: PL

Abstrakty

PL

Materiałem do badań było 65 szczepów Campylobacter jejuni izolowanych w Zakładzie Bakteriologii Państwowego Zakładu Higieny w latach 2003-2005 z próbek kalu dzieci z biegunką. Wśród badanych szczepów 58% stanowiły szczepy oporne na ciprofloksacynę. Przy użyciu metody PCR-RFLP określono częstość występowania mutacji w kodonie 86 genu gyrA kodującym podjednostkę A gyrazy DNA u C. jejuni. U wszystkich szczepów opornych na ciprofloksacynę (MIC dla ciprofloksacyny > 32 µg/ml) wykryto mutację w kodonie 86 genu gyrA, natomiast szczepy wrażliwe na ciprofloksacynę nie posiadały tej mutacji.

EN

The aim of this study was to investigate the mutations in gyrA gene at Thr-86 position in fluoroquinolone resistant C. jejuni clinical isolates (2003-2005). The change of Thr to lie at 86 position is associated with high-level resistance to fluoroquinolone in C. jejuni. Thirty five (58%) of 65 C. jejími strains were found to be resistant to ciprofloxacin using E-test method. PCR-RFLP technique with the Rsal enzyme was used for the identification of mutation in gyrA gene. The primers spanning a part of the fluoroquinolone resistance determining region (QRDR) were designed based on the article of Alon- so et al. and the gyrA sequence of C. jejuni (Gen Bank accession number L04566). One of this primer had mismatch introduced at the second nucleotide from 3 ' end of the primer what gives an artificial Rsal cleavage site. All of the ciprofloxacin-resistant isolates contained a single point mutation in the gyrA gene: the replacement of Thr 86 by lie. The results showed that PCR-RFLP is a rapid and simple method for the detection of the high-level fluoroquinolone resistance in C. jejuni.

Słowa kluczowe

PL

szcepy bakteryjne; Campylobacter jejuni; dzieci; biegunki; probki kalu; oznaczanie; metoda PCR-RFLP; lekoopornosc; ciprofloksacyna; gen gyrA; fluorochinolony; mutacje genowe; kodon 86

• Wydawca: -
• Czasopismo: Medycyna Doświadczalna i Mikrobiologia
• Rocznik: 2005
• Tom: 57
• Numer: 3
• Opis fizyczny: s.295-301,rys.,bibliogr.

Twórcy

autor

Wardak S.

• Panstwowy Zaklad Higieny w Warszawie, ul.Chocimska 24, 00-791 Warszawa

autor

Szych J.

autor

Cieslik A.

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