کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1177540 962026 2006 10 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A 96-well formatted method for exon and exon/intron boundary full sequencing of the CFTR gene
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
A 96-well formatted method for exon and exon/intron boundary full sequencing of the CFTR gene
چکیده انگلیسی

Full genotypic characterization of subjects affected by cystic fibrosis (CF) is essential for the definition of the genotype–phenotype correlation as well as for the enhancement of the diagnostic and prognostic value of the genetic investigation. High-sensitivity diagnostic methods, capable of full scanning of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, are needed to enhance the significance of these genetic assays. A method for extensive sequencing of the CFTR gene was optimized. This method was applied to subjects clinically positive for CF and to controls from the general population of central Italy as well as to a single subject heterozygous for a mild mutation and with an uncertain diagnosis. Some points that are crucial for the optimization of the method emerged: a 96-well format, primer project and purification, and amplicon purification. The optimized method displayed a high degree of diagnostic sensitivity; we identified a subset of 13 CFTR mutations that greatly enhanced the diagnostic sensitivity of common methods of mutational analysis. A novel G1244R disease causing mutation, leading to a CF phenotype with pancreatic sufficiency but early onset of pulmonary involvement, was detected in the subject with an uncertain diagnosis. Some discrepancies between our results and previously published CFTR sequence were found.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 353, Issue 2, 15 June 2006, Pages 226–235
نویسندگان
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