Intracellular killing of bacteria and fungi.

Publisher Summary The measurement of bactericidal activity is considered the essential physiological tool that links the clinical expression of phagocytic defects with their biochemical and molecular etiology. Bactericidal activity measurements are often used as a diagnostic screening test in clinical medicine or as a logical source of reference when the inhibitors or enhancers of metabolic steps that influence bacterial killing in phagocytes are experimented. This chapter describes the intracellular killing of bacteria and fungi. It discusses three assay variants that should be applicable to most clinical or experimental laboratory situations. Killing assay using bacterial colony counts (Method 1) can easily be adapted for any bacterial strain by using modifications proposed by Leijh et al. The assessment of microbial killing by a fluorochrome assay (Method 2) is readily applicable to the study of fungi as well as bacteria and uses minuscule amounts of blood. A novel killing assay using a temperature-sensitive mutant as a target organism (Method 3) developed by Hooke confronts the problem of bacterial growth during the course of the assay.