Gram staining was performed from the blood culture broths that showed visible growth. Evidence for growth was considered based on the appearance of colonies growing on top of red cells, turbidity, gas bubbles, or hemolysis. In addition, blind subculturing was done if a visible growth was not detected within 24 hours. Terminal subculturing was done for bottles that did not show visible growth within 7 days. Based on clues from Gram-staining results, we subcultured organisms on chocolate agar, blood agar, MacConkey agar, and Sabouraud dextrose agar (SDA). Two sets of SDA were inoculated for each sample, one incubated at 37°C and the other at room temperature. We incubated bacterial cultures aerobically at 37°C for up to 24–48 hours and the SDA for 2 weeks.
Gram-positive bacteria were identified using coagulase and catalase tests. Enterobacteriaceae isolates and non-fermentative Gram-negative bacilli were identified using analytical profile index (API20E) and API-NE identification kits (BioMérieux, Lyon, France), respectively. For initial identification of Candida albicans, the germ-tube test was used. The thermotolerance test was employed to differentiate C. albicans from C. dubliniensis. Identification of non-C. albicans at a species level was not possible, due to limitations of diagnostics in our setting.
Gram-positive bacteria were identified using coagulase and catalase tests. Enterobacteriaceae isolates and non-fermentative Gram-negative bacilli were identified using analytical profile index (API20E) and API-NE identification kits (BioMérieux, Lyon, France), respectively. For initial identification of Candida albicans, the germ-tube test was used. The thermotolerance test was employed to differentiate C. albicans from C. dubliniensis. Identification of non-C. albicans at a species level was not possible, due to limitations of diagnostics in our setting.