title: A new assay based on terminal restriction fragment length polymorphism of homocitrate synthase gene fragments for Candida species identification.
abstract: Candida sp. have been responsible for an increasing number of infections, especially in patients with immunodeficiency. Species-specific differentiation of Candida sp. is difficult in routine diagnosis. This identification can have a highly significant association in therapy and prophylaxis. This work has shown a new application of the terminal restriction fragment length polymorphism (t-RFLP) method in the molecular identification of six species of Candida, which are the most common causes of fungal infections. Specific for fungi homocitrate synthase gene was chosen as a molecular target for amplification. The use of three restriction enzymes, DraI, RsaI, and BglII, for amplicon digestion can generate species-specific fluorescence labeled DNA fragment profiles, which can be used to determine the diagnostic algorithm. The designed method can be a cost-efficient high-throughput molecular technique for the identification of six clinically important Candida species.
authors.affiliation: Department of Molecular Biotechnology and Microbiology, Faculty of Chemistry, Gdańsk University of Technology, Narutowicza 11/12, 80-233, Gdańsk, Poland.
authors.forename: Kasjan
authors.initials: K
authors.lastname: Szemiako
authors.affiliation: Department of Therapy Monitoring and Pharmacogenetics, Medical University of Gdańsk, Gdańsk, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Śledzińska
authors.affiliation: Department of Molecular Biotechnology and Microbiology, Faculty of Chemistry, Gdańsk University of Technology, Narutowicza 11/12, 80-233, Gdańsk, Poland. beakrawc@pg.gda.pl.
authors.forename: Beata
authors.initials: B
authors.lastname: Krawczyk
mesh_terms: D002175:Candida / Q000201:enzymology / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal; D007652:Oxo-Acid-Lyases / Q000235:genetics*; D012150:Polymorphism, Restriction Fragment Length; D013045:Species Specificity
chemical_list: D004271:DNA, Fungal; C021365:homocitrate synthase; D007652:Oxo-Acid-Lyases
keywords: Candida; Homocitrate synthase gene; Molecular diagnostic; PCR; t-RFLP