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

作者：Szemiako Kasjan; Sledzinska Anna; Krawczyk Beata^*

来源：Journal of Applied Genetics, 2017, 58(3): 409-414.

DOI：10.1007/s13353-017-0394-5

• Candida
• t-RFLP
• PCR
• Homocitrate synthase gene
• Molecular diagnostic

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.

• 出版日期2017-8