Comet Assay as a technique to evaluate DNA damage in sickle cell anemia patients

IMAGES IN CLINICAL HEMATOLOGY

Comet Assay as a technique to evaluate DNA damage in sickle cell anemia patients

Willian Marcel Barberino; Edis Belini-Junior; Claudia Regina Bonini-Domingos

Laboratory of Hemoglobin and Genetics of Hematological Diseases, Biology Department, Universidade Estadual Paulista "Júlio de Mesquita Filho" - UNESP, São José do Rio Preto, SP, Brazil

^{Corresponding author}

Comet Assay is a technique which can detect single-strand breaks as initial DNA damage.^(1,2) Cells submitted to electrophoresis under alkaline conditions in low melting point agarose gel show DNA damage in a comet-like form when viewed at fluorescence microscopy. The lesion from each cell is quantified according to the comet tail length as class 0, 1, 2 or 3.⁽³⁾ In sickle cell disease, due to constant oxidative stress and membrane lesions, this assay can be useful to detect DNA lesion intensity and medication response. Figure 1A illustrates cell nuclei from a 31-year-old sickle cell disease patient taking hydroxyurea classified as comet class 3. Figure 1B shows the result of a 33-year-old subject without any hemoglobinopathy where no lesions are observed.

Submitted: 12/9/2011

Accepted: 1/9/2012

Conflict-of-interest disclosure: The authors declare no competing financial interest

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• 1. Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res. 1988;175(1):184-91.
• 2. Fairbairn DW, Olive PL, O'Neill KL. The Comet assay: a comprehensive review. Mutat Res. 1995;339(1):37-59.
• 3. Olive PL, Banath JP, Durand RE. Heterogeneity in radiation-induced DNA damage and repair in tumor and normal cells using the "comet" assay. Radiat Res. 1990;122(1):86-94.

Publication Dates

• Publication in this collection
  13 Mar 2012
• Date of issue
  2012