Determination of Ionizing-Radiation Exposure Dose Using Quantitative Real Time- PCR in Blood Samples

الموضوعات

علوم تطبيقية وتكنولوجية

الملخص

There is current worldwide concern regarding the possibility of exposure of masses of human beings to ionizing-radiation resulting from major intentional or accidental incidents, such as nuclear accidents or wars. No such dosimetric data is available on Sudanese subjects. The aim of this work is concerned with obtaining such data using a molecular biological method involving gene expression. In this study four radiation sensitive genes were targeted, namely, CDKN1A, DDB2, Bax and P53. The expression of GAPDH, as a ‘house-keeping’ gene, was normalized between samples so that the relative expression of the target genes could be established. Appropriate primers that anneal to the gene coding sequence were used. Blood samples, collected from healthy female and male Sudanese volunteers (with informed consent), ages 27-35 years, were exposed to 1-5 Gy of γ-rays using a caesium-137 irradiation unit at Egypt’s National Center for Radiation Research and Technology, Cairo. The study covered the effect, on gene expression, of different radiation doses as well as the time lapse during which irradiated blood was kept in culture before analyses (namely, 12, 24, 48 and 72 hours). Standard methods (and kits) were used for extraction of irradiated blood total-RNA. Complementary DNA (cDNA) was synthesized from it using reverse transcriptase. Gene expression was evaluated using Quantitative Real Time Polymerase Chain Reaction (QRT-PCR). The results showed a positive correlation (up-regulation) between expression folding of genes and dose of radiation and this correlation was especially clear until the dose of 3 Gy. This linear relationship could be used in dosimetry, especially with CDKN1A gene. At 4 and 5 Gy doses there was also increased response in gene expression but not as sharp as the former doses and was different from one gene to another. The irradiation. In particular P53 gene proved to be a poor biomarker for radiation dosimetry if blood was analyzed 24 hours, or more, after irradiation. Melting curves showed good purity of the PCR products. It is recommended that, in the unfortunate case of mass exposure of Sudanese populations to ionizing radiation, the method of Real-time Quantitative PCR is well suited to use for the determination of radiation dose-exposure of individual victims through the expression of certain genes, particularly CDKN1A gene. This is valid for radiation-exposure dose up to 5 Gy and a time lapse of 24 hours between irradiation and gene expression analysis of CDKN1A, DDB2 and BAX genes.

روابط وملفات

التعريف والنوع

رقم الوثيقة

04469102-a68b-4b0d-b27a-ab56081237f6

رقم العقد

0

نوع الوسائط

Crawler

نوع المحتوى

الرسائل العلمية

صيغة المصدر

رسائل ماجيستير

نوع الملف

pdf text

أسماء الملفات

71836_1.pdf

بيانات النشر

ترجمة العنوان

استخدام طريقة تفاعل إنزيم بوليمريز التسلسلي (QRT - PCR) لتحديد جرعة تعرض للإشعاع المؤين بتشعيع عينات دم

ألقاب المؤلفين

[{"name_ar":"Mutaman Ali A. Kehial","title_ar":"اشراف","title_en":"Supervision"},{"name_ar":"طارق خالد عبدالله المغربي","title_ar":"اشراف","title_en":"Supervision"},{"name_ar":"مجدي بكر محمود أمين","title_ar":"اشراف","title_en":"Supervision"},{"name_ar":"ميادة الفاتح خضر خلف الله","title_ar":"اعداد","title_en":"Preparation"}]

اللغة

English

المصدر والدورية

اسم المصدر

Determination of Ionizing-Radiation Exposure Dose Using Quantitative Real Time- PCR in Blood Samples

المحتوى والصفحات

عدد الصفحات

0

إشراف وإعداد

الإشراف

Mutaman Ali A. Kehial, طارق خالد عبدالله المغربي, مجدي بكر محمود أمين

الإعداد

ميادة الفاتح خضر خلف الله

الاقتباسات الببليوغرافية