مرکز منطقه ای اطلاع رساني علوم و فناوري -

چكيده لاتين :

Background: Yeast infections are increasing cause of morbidity and mort ality in immunocompromised patients. In order to perform a DNA-based diagnostic test, availability of a rapid and easy-to-perform DNA extraction protocol is essenti al. In the present study we evaluated colony-PCR as the easiest way to amplification of target DNA. Methods: Inste ad of using templates of purified genomic DNA, we performed the PCR dire ctly from yeast colon ies or cultures. Serial cell dilution of three reference yeast strains including Candida albicans, Cryptoco ccus neoformans and Saccharomyces cerevisiae were used for determining the sensitivity of the colony-PCR. A total of one hundred yeast isolates were also tested. All reactions were performed using the universal fung al primers ITS I and ITS4 complementary to the rDNA region . Results: The colony- PCR resulted in a single band (with different sizes) for 10 square 6 cells or more for all reference species. Furthermore 98 out of 100 (98%) of samples showed a relevant single band after PCR. Conclusion: Directly application of the yeast cells obtained from culture colony for PCR reaction is a fast , reliable, cost-effective and simple method for performing any PCR-based protocol including diagnostic tests.