Drug resistance is one of the most significant challenges in the treatment of various types of malignancies, however most of the experimental and clinical data in multidrug resistance (MDR) has been obtained in leukemias. MDR is the term that describes innate or acquired resistance of tumor cells to a wide range of anticancer drugs. As its presence determines treatment outcome in several forms of leukemias, it is imperative that clinical laboratories provide the most useful data on its expression. Here, a brief review is provided on the pathomechanism and diagnostics of MDR. From the diagnostic point of view it is fortunate that MDR proteins display similar effluxing activity towards many dissimilar agents some of which can be used in fluorescent assays. These tests mimic the real clinical problem i.e. the extrusion activity of MDR proteins towards xenobiotics. Thus, we believe that functional assays when carried out in a standardized way and particularly combined with labeling for various surface markers can be recommended as a front-line test in MDR measurement.