Malaria is estimated to cause up to 500 million clinical cases and over one million deaths throughout the world each year. It also continues to be a major problem in Thailand, especially in the areas bordering Cambodia and Myanmar. At BIOTEC, the Protein-Ligand Engineering and Molecular Biology Research Program focuses on basic and medical biotechnology, with emphasis on the rational discovery and development of drugs for tropical diseases, especially malaria.
During the process of antimalarial drug development, the active compounds that have shown good antimalarial activity in vitro must be confirmed for their activity using in vivo model, such as experimental animal host infected with Plasmodium parasites. To avoid the conventional counting method under the microscope, a reliable quantitative protocol for flow cytometric enumeration of infected red blood cells stained with SYBR Green I was developed.
In this study, the scientists use the SYBR Green I, one of the brightest and most sensitive dyes specific to the nucleic acid, to stain parasite-infected blood samples. The infected red blood cells are then suitable for counting by a flow cytometer using the bi-dimensional FL-1530/FL-3620 detection method. The technique is highly accurate at quantifying the parasite load of infected mouse blood, and the dye stays stable for hours after staining. This protocol was validated in an antimalarial assay and the result was comparable to that obtained from conventional microscopic counting.
Posted on 26 June 2012