Koloni lizatların restriction fragment length polymorphism (RELP) analizi yolu ile Candida türlerinin tanımlanması ve Candida albicans suşlarında flukonazol direncinin arbitrarily primed polymerase chain reaction (AP-PCR) ile saptanması

Amaç: Bu çalışmada Candida türlerini PCR (Polymerase chain reaction) ve RFLP (restriction fragment lenght polymorphism) analizi ile getotipik yöntemlerle tanımlamaktır. Candida türlerinin PCR ve RFLP analizi ile tanımlanması, rDNA intergenic spacer (ITS) bölgelerinin büyüklük ve primer yapısal varyasyonlarına dayanır. Yöntem: Beş tür halindeki 44 klinik Candida izolatı çalışmaya dahil edilmiştir. Amplifikasyon ürünlerinin herbiri 3 farklı restriksiyon enzimi Hae III, Dde I ve Bfa I tarafından parçalanmıştır. Sonuçlar: Test edilen bütün izolatlar PCR ve RFLP analizi ile beklenen bant örneklerini vermiştir. Bu çalışmadam elde edilen sonuçlar Candida türlerinin sadece Hae III restriksiyon enzimi kullanılarak Candida albicans ve non-C. albicans suşları olarak ayırılabileceğini ve Bfa I'in ayırt edilen non- C. albicans türlerinin doğruladığını ortaya koymaktadır. Bu tanımlama özellikle fungemi vakalarında oldukça önemlidir. Sonuç: Candida türlerinin ITS bölgesinin amplifıkasyonu ve RFLP analizi ile tanımlanması Candida türlerini tanımlamak için kullanılan zaman alıcı konvansiyonel yöntemlerle karşılaştırıldığında pratik, kısa ve güvenilir bir yöntemdir.

Identification of Candida species in restriction fragment length polymorphism (RELP)analysis of colony lysates

Purpose: The aim of this study was to identify Candida species by using genotypic methods such as PCR (polymerase chain reaction) and RFLP (restriction fragment length polymorphism) analysis. The identification of Candida species by PCR and RFLP analysis was based on the size and primary structural variation of rDNA intergenic spacer regions (ITS). Methods: Forty-four clinical Candida isolates comprising 5 species were included to the study. The amplification products were digested individually with 3 different restriction enzymes Haelll, Ddel and B fal. Results: All the isolates tested yielded the expected band patterns by PCR and RFLP analysis. The results obtained from this study demonstrate that Candida species can be differentiated as Candida albicans and non-C. albicans strains only by using HaeIII restriction enzyme and that Bfal confirms the differentiation of these non-C. albicans species. This differentiation seems to be highly efficient especially in cases of fungemia. Conclusion: The identification of Candida species with the amplification of the ITS region and RFLP analysis is a practical, rapid and reliable method when compared with conventional, time-consuming Candida species identification methods

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