Optimization of Alkaline Single Cell Gel Electropohoresis (Comet Assay) Protocol for Cells from Oral Cavity

Abstract

Comet assays or single cell gel electrophoresis for detection of DNA damage is a test that has been widely utilized to assess the effects of expositions to environmental genotoxicants. The test is also used to evaluate DNA damage related to chronic inflammation or preneoplastic and neoplastic conditions. The cells more frequently assessed in comet assay in humans are the peripheral blood lymphocytes, but there are other cell types that have been considered for that purpose. Among those, buccal cells have received attention for its suitability for comet assay, but there have been Original Research Article

Characterization of the external exposome and its contribution to the clinical respiratory and early biological effects in children: The PROMESA cohort study protocol

“…A buccal mucosa sample and a blood sample obtained by capillary puncture will be collected from all children every six months, for the assessment of early biological effects [alkaline Comet assay [54,55] and micronucleus test [56]]. In addition, Benzo[a]pyrene (BaP) adducts in DNA will be determined as an exposure biomarker [57,58], and the polymorphisms of five genes involved in DNA repair and in the metabolism of genotoxic agents (CYP1A1, GSTM, P53, AHR and H2AX) will be determined as susceptibility biomarkers.…”

DNA damage and shell malformation in Blue Mussel, <i>Mytilus edulis</i>

“…Comet assay is a sensitive, simple, flexible, rapid, and inexpensive technique for detecting DNA damage as DNA strand breaks in an individual cell of almost any eukaryotic organism (Azqueta & Collins, 2013;Cvetković et al, 2018;Tatin et al, 2021). Due to its ability and practical advantages, this assay has already been used as a genotoxicity test for various genotoxicants both direct and indirect-acting (Erraud et al, 2018;González et al, 2018;Gajski et al, 2019;Cordelli et al, 2021;Rajendran et al, 2021) for various aquatic organisms from vertebrate (Žegura & Filipič, 2019;Camargo Filho et al, 2022;Cant et al, 2022;Costa, 2022;Gazo et al, 2022;Mahboob et al, 2022) to invertebrate (Erraud et al, 2018;Khan et al, 2019;Guidi et al, 2021;Cosentino et al, 2022;Ivorra et al, 2022) Mussels (Mytilus edulis and their relatives) are a very common aquatic invertebrate species that have been used as sentinel organisms in aquatic environment monitoring since Goldberg (1975) has proposed the Mussel Watch program. Mussels were thought to fulfil what was required by environmental monitoring principles because of their behaviour as a sessile organism, wide geographical distribution, availability, and resistance to general stress.…”