An Optimized In-House Protocol for Cryptococcus neoformans DNA Extraction from Whole Blood Comparison of Lysis Buffer and Ox-Bile Methods.xlsx

This study aimed to develop an economical in-house method for extracting <i>C. neoformans</i> DNA from whole blood. <i>C. neoformans</i> cells of varying McFarland standards were spiked into expired blood, then lysed using laboratory-prepared lysis buffer and ox-bile solution, followed by organic DNA extraction. Ordinary PCR targeting the CNAG 04922 gene was performed. To determine the limit of detection, serial dilutions of <i>C. neoformans</i> were made, and DNA extraction was performed on other parts cultured on yeast extract peptone dextrose agar to determine colony-forming units (CFU). The lysis buffer method successfully extracted DNA from as low as the average of 62 CFU in 0.9 mL of expired blood with superior quality and high yield compared to ox-bile.