Szemiako Kasjan, Śledzińska Anna, Krawczyk Beata
Department of Molecular Biotechnology and Microbiology, Faculty of Chemistry, Gdańsk University of Technology, Narutowicza 11/12, 80-233, Gdańsk, Poland.
Department of Therapy Monitoring and Pharmacogenetics, Medical University of Gdańsk, Gdańsk, Poland.
J Appl Genet. 2017 Aug;58(3):409-414. doi: 10.1007/s13353-017-0394-5. Epub 2017 Mar 27.
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.
念珠菌属已导致越来越多的感染，尤其是在免疫缺陷患者中。在常规诊断中，念珠菌属的种特异性鉴别很困难。这种鉴定在治疗和预防方面可能具有高度显著的关联。这项工作展示了末端限制性片段长度多态性（t-RFLP）方法在六种念珠菌分子鉴定中的新应用，这六种念珠菌是真菌感染最常见的病因。选择真菌特异性的同柠檬酸合酶基因作为扩增的分子靶点。使用三种限制性内切酶DraI、RsaI和BglII对扩增子进行消化，可以产生种特异性的荧光标记DNA片段图谱，可用于确定诊断算法。所设计的方法可以成为一种经济高效的高通量分子技术，用于鉴定六种临床上重要的念珠菌物种。
