Buccal micronucleus cytome assay: Results of an intra- and inter-laboratory scoring comparison

Claudia Bolognesi, Paola Roggieri, Monica Ropolo, Philip Thomas, Maryam Hor, Michael Fenech, Armen Nersesyan, Siegfried Knasmueller

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The buccal micronucleus cytome (BMCyt) assay is a minimally invasive approach for measuring DNA damage, cell proliferation, cell differentiation and cell death in exfoliated buccal cells. The main limitation for its use is the lack of knowledge about inter- and intra-laboratory variability in scoring micronuclei and other end points included in the cytome approach. In order to identify the main sources of variability across the BMCyt biomarkers, a scoring exercise was carried out between three experienced laboratories using the same set of slides and an identical set of detailed scoring criteria and associated images for the different end points. Single batches of slides were prepared from pooled samples of four groups of subjects characterised by different frequencies of cell types and micronuclei, namely Down syndrome patients, head and neck cancer patients undergoing radiotherapy and two age- and gender-matched control groups. A good agreement among the laboratories in the identification of normal differentiated cells and of micronuclei was obtained. A 3-fold and 20-fold increase in the frequency of micronucleated cells and micronuclei in differentiated cells of Down syndrome patients and in cancer patients, respectively, compared to matched controls, was a consistent result in the three laboratories. The scores of other cell types and nuclear anomalies, such as basal, binucleated, condensed chromatin and karyorrhectic cells showed significant disagreement between and within laboratories indicating that their evaluation using the current visual scoring protocol does not yield robust results for these parameters. The guidelines for BMCyt assay application could be improved by combining the anomalies associated with cell death (condensed chromatin and karyorrhectic cells) in a single category and by defining more stringent criteria in classifying basal cell, binucleated cells and buds.

Original languageEnglish
Pages (from-to)545-555
Number of pages11
JournalMutagenesis
Volume30
Issue number4
DOIs
Publication statusPublished - Jul 1 2015

Fingerprint

Micronucleus Tests
Cheek
Assays
Cell death
Chromatin
Down Syndrome
Radiotherapy
Cell proliferation
Biomarkers
Cell Death
Cells
Head and Neck Neoplasms
DNA
DNA Damage
Cell Differentiation
Research Design
Cell Proliferation
Guidelines
Exercise

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)
  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Bolognesi, C., Roggieri, P., Ropolo, M., Thomas, P., Hor, M., Fenech, M., ... Knasmueller, S. (2015). Buccal micronucleus cytome assay: Results of an intra- and inter-laboratory scoring comparison. Mutagenesis, 30(4), 545-555. https://doi.org/10.1093/mutage/gev017

Buccal micronucleus cytome assay : Results of an intra- and inter-laboratory scoring comparison. / Bolognesi, Claudia; Roggieri, Paola; Ropolo, Monica; Thomas, Philip; Hor, Maryam; Fenech, Michael; Nersesyan, Armen; Knasmueller, Siegfried.

In: Mutagenesis, Vol. 30, No. 4, 01.07.2015, p. 545-555.

Research output: Contribution to journalArticle

Bolognesi, C, Roggieri, P, Ropolo, M, Thomas, P, Hor, M, Fenech, M, Nersesyan, A & Knasmueller, S 2015, 'Buccal micronucleus cytome assay: Results of an intra- and inter-laboratory scoring comparison', Mutagenesis, vol. 30, no. 4, pp. 545-555. https://doi.org/10.1093/mutage/gev017
Bolognesi, Claudia ; Roggieri, Paola ; Ropolo, Monica ; Thomas, Philip ; Hor, Maryam ; Fenech, Michael ; Nersesyan, Armen ; Knasmueller, Siegfried. / Buccal micronucleus cytome assay : Results of an intra- and inter-laboratory scoring comparison. In: Mutagenesis. 2015 ; Vol. 30, No. 4. pp. 545-555.
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