title: Buccal micronucleus cytome assay: results of an intra- and inter-laboratory scoring comparison.
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.
authors: Environmental Carcinogenesis Unit, IRCCS Azienda Ospedaliera Universitaria San Martino-IST Istituto Nazionale Ricerca sul Cancro, Largo Rosanna Benzi 10, Genova 16132, Italy, Commonwealth and Scientific Industrial Research Organization, Genome Health and Personalised Nutrition Laboratory, Food and Nutrition Flagship, Adelaide, South Australia 5000, Australia, Institute of Cancer Research, Department of Medicine I, Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8a, A-1090 Vienna, Austria claudia.bolognesi@hsanmartino.it.
authors: Claudia
authors: C
authors: Bolognesi
authors: Environmental Carcinogenesis Unit, IRCCS Azienda Ospedaliera Universitaria San Martino-IST Istituto Nazionale Ricerca sul Cancro, Largo Rosanna Benzi 10, Genova 16132, Italy, Commonwealth and Scientific Industrial Research Organization, Genome Health and Personalised Nutrition Laboratory, Food and Nutrition Flagship, Adelaide, South Australia 5000, Australia, Institute of Cancer Research, Department of Medicine I, Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8a, A-1090 Vienna, Austria.
authors: Paola
authors: P
authors: Roggieri
authors: Environmental Carcinogenesis Unit, IRCCS Azienda Ospedaliera Universitaria San Martino-IST Istituto Nazionale Ricerca sul Cancro, Largo Rosanna Benzi 10, Genova 16132, Italy, Commonwealth and Scientific Industrial Research Organization, Genome Health and Personalised Nutrition Laboratory, Food and Nutrition Flagship, Adelaide, South Australia 5000, Australia, Institute of Cancer Research, Department of Medicine I, Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8a, A-1090 Vienna, Austria.
authors: Monica
authors: M
authors: Ropolo
authors: Commonwealth and Scientific Industrial Research Organization, Genome Health and Personalised Nutrition Laboratory, Food and Nutrition Flagship, Adelaide, South Australia 5000, Australia.
authors: Philip
authors: P
authors: Thomas
authors: Commonwealth and Scientific Industrial Research Organization, Genome Health and Personalised Nutrition Laboratory, Food and Nutrition Flagship, Adelaide, South Australia 5000, Australia.
authors: Maryam
authors: M
authors: Hor
authors: Commonwealth and Scientific Industrial Research Organization, Genome Health and Personalised Nutrition Laboratory, Food and Nutrition Flagship, Adelaide, South Australia 5000, Australia.
authors: Michael
authors: M
authors: Fenech
authors: Institute of Cancer Research, Department of Medicine I, Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8a, A-1090 Vienna, Austria.
authors: Armen
authors: A
authors: Nersesyan
authors: Institute of Cancer Research, Department of Medicine I, Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8a, A-1090 Vienna, Austria.
authors: Siegfried
authors: S
authors: Knasmueller
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000032:analysis; D016022:Case-Control Studies; D016923:Cell Death; D002454:Cell Differentiation / Q000235:genetics; D002467:Cell Nucleus; D049109:Cell Proliferation; D004249:DNA Damage / Q000235:genetics*; D004314:Down Syndrome / Q000235:genetics / Q000473:pathology*; D005260:Female; D006258:Head and Neck Neoplasms / Q000235:genetics / Q000473:pathology* / Q000532:radiotherapy; D006801:Humans; D008297:Male; D015162:Micronucleus Tests / Q000379:methods* / Q000592:standards*; D008875:Middle Aged; D009061:Mouth Mucosa / Q000166:cytology* / Q000648:ultrastructure*; D015588:Observer Variation; D055815:Young Adult
chemical_list: D015415:Biomarkers