Minimum Information for Reporting on the Comet Assay (MIRCA): recommendations for describing comet assay procedures and results

The comet assay is a widely used test for the detection of DNA damage and repair activity. However, there are interlaboratory differences in reported levels of baseline and induced damage in the same experimental systems. These differences may be attributed to protocol differences, although it is difficult to identify the relevant conditions because detailed comet assay procedures are not always published. Here, we present a Consensus Statement for the Minimum Information for Reporting Comet Assay (MIRCA) providing recommendations for describing comet assay conditions and results. These recommendations differentiate between ‘desirable’ and ‘essential’ information: ‘essential’ information refers to the precise details that are necessary to assess the quality of the experimental work, whereas ‘desirable’ information relates to technical issues that might be encountered when repeating the experiments. Adherence to MIRCA recommendations should ensure that comet assay results can be easily interpreted and independently verified by other researchers.

[1]  Steffen Loft,et al.  Statistical analysis of comet assay results , 2014, Front. Genet..

[2]  J.G.J. Mol,et al.  Determination of phomopsin-A in lupin seeds and lupin-derived products : results of an interlaboratory validation study , 2017 .

[3]  Maria Dusinska,et al.  The comet assay in animal models: From bugs to whales - (Part 2 Vertebrates). , 2019, Mutation research.

[4]  Amaya Azqueta,et al.  Does the duration of lysis affect the sensitivity of the in vitro alkaline comet assay? , 2015, Mutagenesis.

[5]  Maria Dusinska,et al.  On the search for an intelligible comet assay descriptor , 2014, Front. Genet..

[6]  Douglas S. Ehrman,et al.  The Technical Approach , 2012 .

[7]  Gunnar Brunborg,et al.  The comet assay: topical issues. , 2008, Mutagenesis.

[8]  Jason E. Stewart,et al.  Minimum information about a microarray experiment (MIAME)—toward standards for microarray data , 2001, Nature Genetics.

[9]  Amaya Azqueta,et al.  Standardisation of the in vitro comet assay: influence of lysis time and lysis solution composition on the detection of DNA damage induced by X-rays , 2018, Mutagenesis.

[10]  Maria Dusinska,et al.  Technical recommendations to perform the alkaline standard and enzyme-modified comet assay in human biomonitoring studies. , 2019, Mutation research.

[11]  Steffen Loft,et al.  Variation in assessment of oxidatively damaged DNA in mononuclear blood cells by the comet assay with visual scoring. , 2008, Mutagenesis.

[12]  Amaya Azqueta,et al.  The next three decades of the comet assay: a report of the 11th International Comet Assay Workshop. , 2017, Mutagenesis.

[13]  Peter Møller,et al.  Measurement of DNA damage with the comet assay in high-prevalence diseases: current status and future directions. , 2019, Mutagenesis.

[14]  Amaya Azqueta,et al.  The enzyme-modified comet assay: Enzyme incubation step in 2 vs 12-gels/slide systems. , 2019, Mutation research.

[15]  Escodd.  Comparative analysis of baseline 8-oxo-7,8-dihydroguanine in mammalian cell DNA, by different methods in different laboratories: an approach to consensus. , 2002, Carcinogenesis.

[16]  Amaya Azqueta,et al.  Measurement of DNA base and nucleotide excision repair activities in mammalian cells and tissues using the comet assay--a methodological overview. , 2013, DNA repair.

[17]  Takashi Omori,et al.  Statistical issues in the use of the comet assay. , 2008, Mutagenesis.

[18]  David H Phillips,et al.  Inter-laboratory variation in DNA damage using a standard comet assay protocol. , 2012, Mutagenesis.

[19]  V. Beneš,et al.  The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. , 2009, Clinical chemistry.

[20]  P. Olive,et al.  Heterogeneity in Radiation-Induced DNA Damage and Repair in Tumor and Normal Cells Measured Using the “Comet” Assay , 1990 .

[21]  David H Phillips,et al.  Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories. , 2014, Mutagenesis.

[22]  Steffen Loft,et al.  An ECVAG† trial on assessment of oxidative damage to DNA measured by the comet assay , 2009, Mutagenesis.

[23]  Paolo Vineis,et al.  STrengthening the Reporting of OBservational studies in Epidemiology--Molecular Epidemiology (STROBE-ME): an extension of the STROBE statement. , 2011, Mutagenesis.

[24]  Amaya Azqueta,et al.  Towards a more reliable comet assay: optimising agarose concentration, unwinding time and electrophoresis conditions. , 2011, Mutation research.

[25]  João Paulo Teixeira,et al.  Potassium bromate as positive assay control for the Fpg-modified comet assay. , 2020, Mutagenesis.

[26]  John P. A. Ioannidis,et al.  Erratum: STrengthening the reporting of observational studies in epidemiology - Molecular epidemiology (STROBE-ME): An extension of the STROBE statement (Mutagenesis (2012) 27:1 (17-29)) , 2012 .

[27]  Mahsa Karbaschi,et al.  Simplified method for the collection, storage, and comet assay analysis of DNA damage in whole blood. , 2011, Free radical biology & medicine.

[28]  Emilio Rojas,et al.  Epithelial cells as alternative human biomatrices for comet assay , 2014, Front. Genet..

[29]  R E Durand,et al.  Factors influencing DNA migration from individual cells subjected to gel electrophoresis. , 1992, Experimental cell research.

[30]  David H Phillips,et al.  Variation in the measurement of DNA damage by comet assay measured by the ECVAG inter-laboratory validation trial. , 2010, Mutagenesis.

[31]  ESCODD,et al.  Measurement of DNA oxidation in human cells by chromatographic and enzymic methods , 2003 .

[32]  David H Phillips,et al.  An ECVAG inter-laboratory validation study of the comet assay: inter-laboratory and intra-laboratory variations of DNA strand breaks and FPG-sensitive sites in human mononuclear cells. , 2013, Mutagenesis.

[33]  Peter Møller,et al.  DNA repair as a human biomonitoring tool: Comet assay approaches. , 2019, Mutation research.

[34]  Paolo Vineis,et al.  STrengthening the Reporting of OBservational studies in Epidemiology - Molecular Epidemiology (STROBE-ME): an extension of the STROBE statement. , 2011, Preventive medicine.

[35]  Peter Møller,et al.  Assessment and reduction of comet assay variation in relation to DNA damage: studies from the European Comet Assay Validation Group. , 2010, Mutagenesis.

[36]  Peter Møller,et al.  Application of the comet assay in human biomonitoring: An hCOMET perspective. , 2020, Mutation research.

[37]  Gunnar Brunborg,et al.  Electrophoresis in the Comet Assay , 2018, Electrophoresis - Life Sciences Practical Applications.

[38]  Steffen Loft,et al.  DNA-repair measurements by use of the modified comet assay: an inter-laboratory comparison within the European Comet Assay Validation Group (ECVAG). , 2013, Mutation research.

[39]  Nan Zhang,et al.  Identification of Fatty Acid Desaturase 6 in Golden Pompano Trachinotus Ovatus (Linnaeus 1758) and Its Regulation by the PPARαb Transcription Factor , 2018, International journal of molecular sciences.

[40]  Lennart Möller,et al.  The effects on DNA migration of altering parameters in the comet assay protocol such as agarose density, electrophoresis conditions and durations of the enzyme or the alkaline treatments. , 2011, Mutagenesis.

[41]  J. Ioannidis,et al.  STrengthening the Reporting of OBservational studies in Epidemiology – Molecular Epidemiology (STROBE-ME): An Extension of the STROBE Statement , 2011, PLoS medicine.

[42]  Steffen Loft,et al.  Harmonising measurements of 8-oxo-7,8-dihydro-2′-deoxyguanosine in cellular DNA and urine , 2012, Free radical research.

[43]  Stuart Moss,et al.  Current Status and Future Directions , 2013 .

[44]  Andrew Collins,et al.  Establishing the background level of base oxidation in human lymphocyte DNA: results of an interlaboratory validation study , 2005, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[45]  Steffen Loft,et al.  Searching for assay controls for the Fpg- and hOGG1-modified comet assay , 2018, Mutagenesis.

[46]  A. Azqueta,et al.  Applying the comet assay to fresh vs frozen animal solid tissues: A technical approach. , 2019, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[47]  E. Khizhnyak,et al.  Some causes of inter-laboratory variation in the results of comet assay. , 2014, Mutation research. Genetic toxicology and environmental mutagenesis.

[48]  Mahsa Karbaschi,et al.  Evaluation of the Major Steps in the Conventional Protocol for the Alkaline Comet Assay , 2019, International journal of molecular sciences.

[49]  Maria Dusinska,et al.  Measurement of DNA oxidation in human cells by chromatographic and enzymic methods. , 2003, Free radical biology & medicine.

[50]  Maria Dusinska,et al.  The comet assay in animal models: From bugs to whales - (Part 1 Invertebrates). , 2019, Mutation research.