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A new assay based on terminal restriction fragment length polymorphism of homocitrate synthase gene fragments for Candida species identification

Authors
  • Szemiako, Kasjan1
  • Śledzińska, Anna2
  • Krawczyk, Beata1
  • 1 Gdańsk University of Technology, Department of Molecular Biotechnology and Microbiology, Faculty of Chemistry, Narutowicza 11/12, Gdańsk, 80-233, Poland , Gdańsk (Poland)
  • 2 Medical University of Gdańsk, Department of Therapy Monitoring and Pharmacogenetics, Gdańsk, Poland , Gdańsk (Poland)
Type
Published Article
Journal
Journal of applied genetics
Publication Date
Mar 27, 2017
Volume
58
Issue
3
Pages
409–414
Identifiers
DOI: 10.1007/s13353-017-0394-5
Source
Springer Nature
Keywords
License
Green

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.

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