The single cell gel electrophoresis assay (comet assay): A European review

McKelvey‐Martin, Valerie J.; Green, M.H.L.; Schmezer, Peter; Pool-Zobel, B.L.; Méo, M.P. De; Collins, Andrew

doi:10.1016/0027-5107(93)90207-v

Cited by 809 publications

(334 citation statements)

References 28 publications

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“…Presumably it is undetected by nick translation because the assay Table I DNA requires clean 3'-OH ends for polymerase activity. Hydrogen peroxide is a potent DNA strand-breaking agent, at very low doses (C 100 PM) in the comet assay [30], which should detect any type of nicking of supercoiled domains in the residual nuclear structure of the lysed cell [31]. "Dirty' strand breaks may indeed be detected more readily in the comet assay because they are less rapidly repaired.…”

Section: Discussionmentioning

confidence: 99%

Endogenous nitric oxide induced by interleukin‐1β in rat islets of Langerhans and HIT‐T15 cells causes significant DNA damage as measured by the ‘comet’ assay

et al. 1993

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We have used the comet assay (single cell gel electrophoresis) to measure nitric oxide-induced DNA damage in rat islets of Langerhans and insulin-containing HIT-T15 cells. Damage was induced following treatment with the nitric oxide donor SIN-l, which also releases superoxide, but was not reduced by exogenous superoxide dismutase, suggesting that nitric oxide itself, rather than superoxide or peroxynitrite may be the active species. The DNA damaging effect of nitric oxide was easily detectable at the earliest time point tested (15 min). Damage also resulted following induction of nitric oxide synthase by the cytokine interleukin-I/? in both islets and HIT-T15 cells and was prevented by replacing the substrate, arginine, with nitromonomethyl arginine. Thus intracellular levels of nitric oxide generated by interleukin-l/l-induced nitric oxide synthase were sufficient to cause DNA damage in islet cells and HIT-T15 cells.

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Section: Discussionmentioning

confidence: 99%

Endogenous nitric oxide induced by interleukin‐1β in rat islets of Langerhans and HIT‐T15 cells causes significant DNA damage as measured by the ‘comet’ assay

et al. 1993

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“…Furthermore, together with the assessment of in vivo genotoxicity of chemicals in multiple organs 31 or after environmental and occupational exposure, 32,33 it is an ideal tool for assessing variability in the response of different cell types in vitro. 34,35 The terminal deoxynucleotidyl transferasemediated dUTP nick-end labeling (TUNEL) assay has also proved to be a sensitive and reliable method for measuring DNA strand breaks. However, the detection of highly damaged cells by the Comet assay occurred earlier than the detection of DNA modifications by the TUNEL assay 36 and more sensitively.…”

Section: Introductionmentioning

confidence: 99%

Amifostine (WR-2721) selective protection against melphalan genotoxicity

et al. 2000

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“…Una de las técnicas utilizada para esta vigilancia es el ensayo del cometa o la electroforesis alcalina de células individuales, que permite evaluar el daño causado en el material genético por diferentes agentes químicos y físicos (21)(22)(23). Tiene amplias aplicaciones en toxicología genética, en ensayos de genotoxicidad in vitro (24)(25)(26)(27)(28)(29) e in vivo (30)(31)(32), en vigilancia biológica ambiental y en vigilancia de poblaciones humanas expuestas a diversas sustancias químicas mutagénicas (33)(34)(35)(36)(37)(38)(39)(40).…”

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Evaluación del daño en el ADN y vigilancia biológica de la exposición laboral a solventes orgánicos, 2006

et al. 2008

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Introducción. La exposición a solventes es uno de los mayores riesgos potenciales para millones de trabajadores en el mundo; los solventes generan contaminación ambiental y desencadenan problemas de salud pública. Objetivo. Determinar los niveles de los metabolitos benceno, tolueno y xileno, los polimorfismos de las enzimas CYP2E1, GSTM1, GSTT1 y el daño del ADN mediante el ensayo del cometa. Materiales y métodos. Se llevó a cabo un estudio descriptivo de corte transversal para la determinación de polimorfismos genéticos y prueba del cometa en 90 trabajadores pertenecientes a cinco empresas. Se aplicó una encuesta, se tomaron muestras de sangre y de orina, se midieron las concentraciones de fenol, ácido hipúrico orto y meta-metilhipúrico. Se hizo el análisis estadístico y se exploraron posibles asociaciones. Resultados. El 34,4% eran trabajadores con exposición directa a solventes. En este grupo se evidenciaron concentraciones superiores a los límites permisibles en 3,3% para fenol, en 6,6% para ácido hipúrico, en 3,3% para ácido orto-metilhipúrico y en 36,7% para ácido metametilhipúrico, mayor longitud promedio de la cola del cometa (19,5 µm) y un incremento del porcentaje de células con daño medio (19,0%) (p=0,0007). El porcentaje de individuos expuestos con genotipos ausentes para las enzimas GSTT1 y GSTM1 fue de 46,7% y de 56,8%, respectivamente. Conclusión. El uso de biomarcadores de exposición, efecto y susceptibilidad, se ha convertido en una herramienta fundamental para la evaluación del riesgo asociado con la exposición a agentes tóxicos.Palabras clave: daño del ADN/genética, exposición profesional, solventes/toxicidad, salud pública DNA damage assessment and biological monitoring of occupational exposure to organic solvents, 2006Introduction. Exposure to solvents is one of the highest potential risks for millions of workers in the world; they can generate substantial environmental pollution leading to outbreaks of public health problems. Objective. Blood levels were determined for metabolites of benzene, toluene and xylene, and polymorphisms for enzymes CYP2E1, GSTM1, GSTT1 were characterized. Damage to DNA was assessed by the comet assay for exposure to organic solvents. Materials and methods. A cross-sectional study was undertaken in 90 employees from 5 different companies. A survey form was administered; blood was sampled to detect the genetic polymorphisms and to apply the comet assay (single cell gel electrophoresis) to detect DNA fragmentation. The concentrations of phenol, ortho and meta methylhippuric acids were measured in urine. Statistical analyses explored the possible associations. Results. The percentage of workers directly exposed to solvents was 34.4%. In this group, the evidence indicated concentrations higher than the permitted limits: 3.3% for phenol, 6.6% for

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The single cell gel electrophoresis assay (comet assay): A European review

Cited by 809 publications

References 28 publications

Endogenous nitric oxide induced by interleukin‐1β in rat islets of Langerhans and HIT‐T15 cells causes significant DNA damage as measured by the ‘comet’ assay

Endogenous nitric oxide induced by interleukin‐1β in rat islets of Langerhans and HIT‐T15 cells causes significant DNA damage as measured by the ‘comet’ assay

Amifostine (WR-2721) selective protection against melphalan genotoxicity

Evaluación del daño en el ADN y vigilancia biológica de la exposición laboral a solventes orgánicos, 2006

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