Effect Of Candida Albicans Cell Concentration On Dna Isolation Results Using Modified Alkaline Lysis Method

Candida albicans is one of the leading causes of morbidity and mortality in various countries, with the prevalence of infection reaching 17,640 cases with an average incidence of 1.5 million people, causing more than one billion people, and the adverse effects of fungal infections can threaten >1.6 million lives. This study aimed to determine the effect of Candida albicans cell concentration on DNA isolation results. The method used was the modified Alkaline lysis using cell cultures with concentrations of 10⁸, 10⁹, and 10⁰ cells/mL. The results showed that the growth curve of the sample showed an increase in cell concentration directly proportional to the absorbance of the UV-Vis spectrophotometer results. Cell suspensions in SDB media were incubated for 2-3 days, then the absorbance was measured, resulting in a value of 5.307 nm after multiplying the dilution factor. The results of Candida albicans DNA isolation were visualized using 2% agarose gel electrophoresis, with no DNA bands found in all samples and negative results. In contrast, smear bands appeared in positive controls. Analysis using a Nanodrop spectrophotometer showed the purity of DNA isolates at concentrations of 10⁸ and 10⁹ < 1.8, while at 10¹⁰ it was 1.82. The average DNA concentration obtained was < 100 ng/µL. Based on the results of statistical tests, it can be concluded that the p-value <0.05 indicates the effect of cell concentration on the results of DNA isolation using the modified Alkaline lysis method. Keywords: Cell concentration, Candida albicans, Alkaline lysis, DNA isolation