Performance and consistency of a fluorescence-based high-throughput screening assay for use in Babesia drug screening in mice.

ABSTRACT: In this study, we evaluated the validity of a fluorescence-based assay using SYBR Green I (SG I) stain for screening antibabesial compounds against B. microti in mice. Two different hematocrits (HCTs; 2.5% and 5%) were used. Correlating relative fluorescence units (RFUs) with parasitemia showed significant linear relationships with R² values of 0.97 and 0.99 at HCTs of 2.5% and 5%, respectively. Meanwhile, the Z' factors in a high-throughput screening (HTS) assay were within the permissible limit (≥0.5) at 2.5% HCT and lower than this value at 5% HCT. Taken together, the highest signal-to-noise (S/N) ratios were obtained at 2.5% HCT; therefore, we concluded that 2.5% was the best HCT for applying fluorescence assay in antibabesial drug screening in mice. Additionally, positive c

SUBMITTER: Rizk MA 

PROVIDER: S-EPMC5643553 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature