Phospho-SMC1 in-Cell ELISA based Detection of Ataxia Telangiectasia | ||
| Journal of Pediatric Perspectives | ||
| مقاله 1، دوره 4، شماره 12 - شماره پیاپی 36، اسفند 2016، صفحه 3957-3967 اصل مقاله (467.51 K) | ||
| شناسه دیجیتال (DOI): 10.22038/ijp.2016.7751 | ||
| نویسندگان | ||
| Majid Zaki dizaji1؛ Nima Rezaei2؛ Marjan Yaghmaie3؛ Mehdi Yaseri4؛ Seyed Javad Sayedi5؛ Gholamreza Azizi6؛ Asghar Aghamohammadi* 2؛ Seyed Mohammad Akrami* 7 | ||
| 1Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences | ||
| 2Research Center for Immunodeficiencies, Children's Medical Center, Tehran University of Medical Science, Tehran, Iran | ||
| 3Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences | ||
| 4Department of Epidemiology and Biostatistics, Tehran University of Medical Sciences | ||
| 5Department of Pediatrics, School of Medicine, Mashhad University of Medical sciences, Mashhad, Iran | ||
| 6Department of Laboratory Medicine, Imam Hassan Mojtaba Hospital, Alborz University of Medical Sciences, Karaj, Iran. | ||
| 7Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran | ||
| چکیده | ||
| Background Ataxia telangiectasia (A-T) is a common genetically inherited cause of early childhood-onset ataxia. The infrequency of this disease, vast phenotype variation, disorders with features similar to those of A-T, and lack of definite laboratory test, make diagnosis difficult. In addition, there is no rapid reliable laboratory method for identifying A-T heterozygotes, who susceptible to ionizing radiation (IR), atherosclerosis, diabetes, and cancers. We used SMC1pSer966 (pSMC1) in-cell colorimetric ELISA to diagnosis and screen in A-T families. Materials and Methods: With informed consent, 2cc peripheral blood was collected from the 15 A-T patients, their parents, and 24 healthy controls with no family history of malignancy, diabetes, and atherosclerosis. Extracted peripheral blood mononuclear cells (PBMCs) were cultured in poly-L-Lysine treated 96-well plate with density of 70,000 cells per well. SMC1 phosphorylation was evaluated with cell-based ELISA kit 1 hour after 5 Gy IR and the pSMC1data normalized with Glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Results: SMC1 phosphorylation was significantly low in A-T`s PBMC (mean + standard deviation [SD]: 0.075 + 0.034) in comparison to carriers (mean + SD: 0.190 + 0.060) and healthy controls (mean + SD: 0.312 +0.081), but unluckily could only discriminate A-T patients (Area Under the Curve -receiver operating characteristic [AUC-ROC]: 1.00, 1.00-1.00). This method in spite of rapidness and simplicity showed poor imprecision (22.49% coefficient of variation [CV] for intraday imprecision). Conclusion: It seems pSMC1 assessment by in-cell ELISA can be used for detection of A-T patients, but it may not sensitive enough for identification of carriers. This ELISA test is very simple, rapid, and requires less than 2cc blood. Thus it may be proposed for the early differential diagnosis of A-T as an alternative method. | ||
| کلیدواژهها | ||
| Ataxia telangiectasia؛ Children؛ ELISA؛ PBMC؛ SMC1 | ||
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