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

Möller, Peter; Azqueta, Amaya; Boutet-Robinet, Élisa; Koppen, Gudrun; Bonassi, Stefano; Milić, Mirta; Gajski, Goran; Costa, Solange; Teixeira, João Paulo; Pereira, Cristiana; Dušinská, Maria; Godschalk, Roger; Brunborg, Gunnar; Gützkow, Kristine B.; Giovannelli, Lisa; Cooke, Marcus S.; Richling, Elke; Laffón, Blanca; Valdiglesias, Vanessa; Başaran, Nurşen; Bò, Cristian Del; Žegura, Bojana; Novak, Matjaž; Stopper, Helga; Vodička, Pavel; Vodenková, Soňa; Andrade, Vanessa Moraes de; Šrámková, Monika; Gábelová, Alena; Collins, Andrew; Langie, Sabine A. S.

doi:10.1038/s41596-020-0398-1

Cited by 229 publications

(118 citation statements)

References 44 publications

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“…This potential problem seems to be mitigated by the use of analysing software, which all of the papers described below used. In 2020, Moller et al published detailed recommendations for reporting comet assay results [46]. For the most objective analysis of the data, the authors suggest the use of calibration curves and conversion of results to relative lesion frequency compared to unaltered nucleotides.…”

Section: Alkaline Comet Assaymentioning

confidence: 99%

Use of Biological Dosimetry for Monitoring Medical Workers Occupationally Exposed to Ionizing Radiation

Piotrowski

Dawid²,

Kulcenty

et al. 2021

Radiation

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Medical workers are the largest group exposed to man-made sources of ionizing radiation. The annual doses received by medical workers have decreased over the last several decades, however for some applications, like fluoroscopically guided procedures, the occupational doses still remain relatively high. Studies show that for some procedures the operator and staff still use insufficient protective and dosimetric equipment, which might cause an underestimation of medical exposures. Physical dosimetry methods are a staple for estimating occupational exposures, although due to the inconsistent use of protection measures, an alternative method such as biological dosimetry might complement the physical methods to achieve a more complete picture. Such methods were used to detect exposures to doses as low as 0.1 mSv/year, and could be useful for a more accurate assessment of genotoxic effects of ionizing radiation in medical workers. Biological dosimetry is usually based on the measurement of the effects present in peripheral blood lymphocytes. Although some methods, such as chromosome aberration scoring or micronucleus assay, show promising results, currently there is no one method recognized as most suitable for dosimetric application in the case of chronic, low-dose exposures. In this review we decided to evaluate different methods used for biological dosimetry in assessment of occupational exposures of medical workers.

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Section: Alkaline Comet Assaymentioning

confidence: 99%

Use of Biological Dosimetry for Monitoring Medical Workers Occupationally Exposed to Ionizing Radiation

Piotrowski

Dawid²,

Kulcenty

et al. 2021

Radiation

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“…Minimum Information for Reporting Comet Assay procedures and results (MIRCA) recommendations were followed in this manuscript (Møller et al 2020a ). After MMS, Act-D and UV treatments, primary DNA damage was assessed by the standard alkaline comet assay.…”

Section: Methodsmentioning

confidence: 99%

Salivary leucocytes as suitable biomatrix for the comet assay in human biomonitoring studies

et al. 2021

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Peripheral blood leucocytes (PBL) have been traditionally used to investigate DNA damage by the comet assay in population studies, but validating alternative non-invasive samples would expand the application of this assay in human biomonitoring. The objectives of this study were (i) to test the validity of salivary leucocytes as a proper biomatrix for the comet assay, (ii) to evaluate the ability of this approach to detect different types of primary and oxidative DNA damage, and (iii) to determine whether frozen salivary leucocytes are still suitable for displaying those types of DNA damage. Fresh and frozen leucocytes isolated from saliva samples (six healthy non-smoking volunteers), were exposed to four genotoxic agents inducing different types of DNA damage, both primary (methyl methanesulfonate, actinomycin-D, ultraviolet radiation) and oxidative (potassium bromate), and standard or enzyme-modified comet assay was conducted. Results were compared with those obtained from PBL. Cells exposed to the four genotoxic agents showed dose-dependent increases of primary and oxidative DNA damage, demonstrating the suitability of all these samples to detect genetic damage from different origin. When comparing baseline levels of DNA damage, just a slight significant increase in primary DNA damage was observed in frozen salivary leucocytes regarding the other biomatrices, but similar results were obtained regarding sensitivity to DNA damage induction by all agents tested. This study demonstrates that salivary leucocytes can be employed in comet assay as an alternative or complement to blood samples. Frozen salivary leucocytes were proved to be a very convenient sample in large biomonitoring studies.

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“…The work of hCOMET has led to technical recommendations of the enzyme-modified comet assay [ 292 ], and detailed reviews on the use of the comet assay for assessment of oxidatively damaged DNA and BER activity in human biomonitoring studies [ 293 , 294 ]. A rather striking outcome is also the Minimum Information for Reporting Comet Assay (MIRCA) recommendations, which has been fostered to increase the quality and impact of comet assay results, including oxidatively damaged DNA [ 295 ]. Alongside this effort, there has been a gentle push for comet assay researchers to use and report assay controls in their experimental procedure as only about 20% of published articles mention the use, or show results from assay controls [ 296 , 297 ].…”

Section: The Futurementioning

confidence: 99%

Biomarkers of nucleic acid oxidation – A summary state-of-the-art

Chao

Evans

et al. 2021

Redox Biology

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Oxidatively generated damage to DNA has been implicated in the pathogenesis of a wide variety of diseases. Increasingly, interest is also focusing upon the effects of damage to the other nucleic acids, RNA and the (2′-deoxy-)ribonucleotide pools, and evidence is growing that these too may have an important role in disease. LC-MS/MS has the ability to provide absolute quantification of specific biomarkers, such as 8-oxo-7,8-dihydro-2′-deoxyGuo (8-oxodG), in both nuclear and mitochondrial DNA, and 8-oxoGuo in RNA. However, significant quantities of tissue are needed, limiting its use in human biomonitoring studies. In contrast, the comet assay requires much less material, and as little as 5 μL of blood may be used, offering a minimally invasive means of assessing oxidative stress in vivo , but this is restricted to nuclear DNA damage only. Urine is an ideal matrix in which to non-invasively study nucleic acid-derived biomarkers of oxidative stress, and considerable progress has been made towards robustly validating these measurements, not least through the efforts of the European Standards Committee on Urinary (DNA) Lesion Analysis. For urine, LC-MS/MS is considered the gold standard approach, and although there have been improvements to the ELISA methodology, this is largely limited to 8-oxodG. Emerging DNA adductomics approaches, which either comprehensively assess the totality of adducts in DNA, or map DNA damage across the nuclear and mitochondrial genomes, offer the potential to considerably advance our understanding of the mechanistic role of oxidatively damaged nucleic acids in disease.

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

Cited by 229 publications

References 44 publications

Use of Biological Dosimetry for Monitoring Medical Workers Occupationally Exposed to Ionizing Radiation

Use of Biological Dosimetry for Monitoring Medical Workers Occupationally Exposed to Ionizing Radiation

Salivary leucocytes as suitable biomatrix for the comet assay in human biomonitoring studies

Biomarkers of nucleic acid oxidation – A summary state-of-the-art

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