[The comet assay: principles of the method and clinical utilization].
BACKGROUND
Single cell gel electrophoresis or comet assay is used at present in the world to study of DNA damage, DNA repair and apoptosis. The aim of the work is to introduce the principle of comet assay to the medical community and to give a basic survey of its possible clinical applications. The article includes our first experience with this method in detection of apoptosis in bone marrow cells of patients with myelodysplastic syndrome (MDS).


RESULTS
The whole bone marrow aspirates from 6 patients with MDS and 7 control persons were processed by alkaline version of comet assay and the degree of DNA fragmentation in individual cells was quantified using Image Analysis System. In comparison with controls, the patients with diagnosis RA and RARS exhibited in bone marrow significantly elevated number of cells with high level of DNA breaks, reflecting most probably the apoptotic cleavage of DNA. In contrast, the patient in proliferative stage of the disease (MDS-CMML-->AML) exhibited decreased frequency of apoptotic cells, well below the control level.


CONCLUSIONS
Our results correspond with the data published on the occurrence of apoptosis in particular types of MDS. Comet assay represents a simple and cheap technique, applicable in clinical hematology to specify the diagnosis, to monitor the disease progress and efficacy of therapy not only in patients with MDS but also in other diseases resulting from an imbalance between proliferation and apoptosis.
Novotná
B
B
Ustav experimentální medicíny AV CR, Praha. novotna@biomed.cas.cz
Neuwirtová
R
R
Blazková
V
V
Sisková
M
M
D000328:Adult; D000368:Aged; D017209:Apoptosis; D001853:Bone Marrow / Q000473:pathology; D020552:Comet Assay / Q000379:methods; D053938:DNA Fragmentation; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009190:Myelodysplastic Syndromes / Q000473:pathology