Sickle Confirm is an inexpensive screening and confirmatory method developed in our lab for use in underdeveloped countries. The sensitivity and specificity to distinguish sickle cell zygosity is 80%, and we have experienced manufacturing issues in countries without climate control. The purpose of this study is to optimize Sickle Confirm to improve sensitivity and specificity and improve manufacturing in non-climate controlled countries. The hemolyzing agent, saponin, has shown changes in physical characteristics during manufacturing, affecting Sickle Confirm performance. To optimize the manufacturing conditions, saponin stability was observed at room temperature, high temperature, and high humidity in five different containers (white glass, brown glass, white plastic, brown plastic, rubber sealed). Only humidity affected the physical form of saponin. Evaluation of saponin concentration needed to achieve complete hemolysis and the currently used Blood Volume Table was performed to improve the sensitivity and specificity of Sickle Confirm. Fresh powder saponin and old clumped saponin (induced by exposure to humidity) were compared and complete hemolysis was measured. The current recipe requires 0.1% of fresh powder saponin (control), whereas the minimum concentration needed to achieve complete hemolysis for old clumped saponin was 0.5%. The current Blood Volume Table was evaluated to standardize blood inoculum based on patient hematocrit. The hematocrit of AA blood samples was adjusted in 5% increments between 10% and 55%, and varying blood volumes were added to 4 mL distilled H2O to reach an absorbance of 1.350. Adjustments were made to the Blood Volume Table for future Sickle Confirm use.

Dr. Tim Randolph is extraordinarily compassionate and knowledgeable. He truly cares about the success of each and every one of his students and researchers. Dr. Randolph provided constant guidance and support to Kaylie Nichols throughout her research project. Without him, she would have never found the passion for research and hematology that she has today. His dedication to assisting Kaylie over the past three years and helping in the creation of her research presentation is appreciable beyond words."