Recommendations for conducting the in vivo alkaline Comet assay

Abstract

The in vivo alkaline single cell gel electrophoresis assay, hereafter the Comet assay, can be used to investigate the genotoxicity of industrial chemicals, biocides, agrochemicals and pharmaceuticals. The major advantages of this assay include the relative ease of application to any tissue of interest, the detection of multiple classes of DNA damage and the generation of data at the level of the single cell. These features give the Comet assay potential advantages over other in vivo test methods, which are limited largely to proliferating cells and/or a single tissue. The Comet assay has demonstrated its reliability in many testing circumstances and is, in general, considered to be acceptable for regulatory purposes. However, despite the considerable data published on the in vivo Comet assay and the general agreement within the international scientific community over many protocol-related issues, it was felt that a document giving detailed practical guidance on the protocol required for regulatory acceptance of the assay was required. In a recent meeting held in conjunction with the 4th International Comet Assay Workshop (Ulm, Germany, 22-25 July 2001) an expert panel reviewed existing data and recent developments of the Comet assay with a view to developing such a document. This paper is intended to act as an update to the more general guidelines which were published as a result of the International Workshop on Genotoxicity Test Procedures. The recommendations are also seen as a major step towards gaining more formal regulatory acceptance of the Comet assay.

Neurobehavioral and genotoxic parameters of antipsychotic agent aripiprazole in mice

“…The comet assay was used to detect recent DNA damage, such as single and double strand breaks, alkali-labile sites, DNA-DNA and DNA-protein crosslinks [26] . Three hours after the last administration in subchronic treatment, blood and brain samples were collected from the same animals that had been tested in the open-field task to conduct the comet assay.…”

“…Class 1 damage was the most frequent among the aripiprazole-damaged cells, which is considered a reparable minimal damage ( Figure 5). The micronucleus test was used to detect clastogenic/ aneugenic activities, which leads to an increasing frequency of micronuclei, and suggests mutagenic effects at the chromosomal level [15,26] . Aripiprazole has shown positive results in mutagenicity assays [27] .…”

Avaliação da exposição ocupacional ao benzeno em trabalhadores frentistas e analistas de combustíveis utilizando o Teste Cometa como biomarcador de genotoxicidade

“…Para realização do Teste Cometa foi utilizado como amostra biológica o sangue periférico. A metodologia utilizada foi modificada a partir de procedimentos previamente descritos por Rojas et al 23 , Pitozzi et al 24 , Hartman et al 25 , de Tice et al 26 e Azqueta et al 27 . O uso de controles na realização desse procedimento analí-tico permite verificar se o teste foi satisfatório, pois muitas substâncias podem provocar danos ao DNA e, assim, o teste pode funcionar como controle positivo.…”

Potential chemoprotective effects of green propolis, L-lysine and celecoxib on bone marrow cells and peripheral blood lymphocytes of Wistar rats subjected to bladder chemical carcinogenesis

“…The experiments were carried out according to Hartmann et al 12 . The survivors at 40 weeks were used for each experimental group, where slides were prepared from each sample and 100 nuclei/animal were evaluated per slide.…”