A rapid flow cytometry test based on histone H2AX phosphorylation for the sensitive and specific diagnosis of ataxia telangiectasia
| Autor: | Simona Cavalieri, Erica Lantelme, Umberto Ricardi, Luca Orlando, Alfredo Brusco, Paola Porcedda, Dario Gregori, Claudia Giachino, Valentina Turinetto, Antonio Amoroso |
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| Rok vydání: | 2008 |
| Předmět: |
flow citometry
Histology Ataxia Immunoblotting Cell Cycle Proteins Ataxia Telangiectasia Mutated Proteins Protein Serine-Threonine Kinases Biology Sensitivity and Specificity Peripheral blood mononuclear cell Pathology and Forensic Medicine Flow cytometry Diagnosis Differential Histones Ataxia Telangiectasia medicine Humans H2AX Phosphorylation Oculomotor apraxia Phosphorylated Histone H2AX medicine.diagnostic_test Tumor Suppressor Proteins Reproducibility of Results Cell Biology Reference Standards Flow Cytometry medicine.disease Molecular biology DNA-Binding Proteins ATM Ataxia-telangiectasia Leukocytes Mononuclear medicine.symptom Nijmegen breakage syndrome |
| Zdroj: | Cytometry Part A. :508-516 |
| ISSN: | 1552-4930 1552-4922 |
| DOI: | 10.1002/cyto.a.20566 |
| Popis: | Ataxia telangiectasia (A-T) is a progressive neurodegenerative disease with onset in early childhood, caused by mutations in the ATM (ataxia-telangiectasia mutated) gene. Diagnosis relies on laboratory tests showing high levels of serum alphafetoprotein, cell sensitivity to ionizing radiation (IR) and absence or reduced levels of ATM protein. Many tests, however, are not sufficiently sensitive or specific for A-T, have long turnaround times, or require large blood samples. This prompted us to develop a new flow cytometry method for the diagnosis of A-T based on the measurement of histone H2AX phosphorylation. We established normal ranges of histone H2AX phosphorylation after 2 Gy IR by testing T-cell lines, lymphoblastoid cell lines (LCLs) and/or peripheral blood mononuclear cells (PBMCs) or both from 20 genetically proven A-T and 46 control donors. To further evaluate the specificity and sensitivity of the test, we analyzed cells from 19 patients suspected of having A-T, and from one Friedreich Ataxia, one Ataxia with Oculomotor Apraxia type 2, and one Nijmegen Breakage Syndrome patients. Phosphorylated histone H2AX mean fluorescence intensity of irradiated A-T cells was significantly lower than that of healthy donors. The intrastaining, intraassay, and interassay imprecisions were |
| Databáze: | OpenAIRE |
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