Škrlec Ivana, Barišić Karmela, Wagner Jasenka
Faculty of Dental Medicine and Health, J. J. Strossmayer University of Osijek, Croatia
Department of Medical Biology and Genetics, Faculty of Medicine, J. J. Strossmayer University of Osijek, Croatia.
Ann Clin Lab Sci. 2018 Nov;48(6):810-813.
The objectives of this study were to validate the direct triplet-primed PCR method (dTP-PCR) for determination of dynamic mutations in the gene, and to compare the results of the dTP-PCR method and Southern blot analysis. The number of CGG repeats in the gene was determined by the direct triplet-primed PCR method and by melting curve analysis. The cut-off temperature between normal and permutations of the CGG repeats was determined using control samples with a known number of CGG repeats. All patients are classified into four categories based on the DNA melting curve. The clinical performance of the assay was established by 40 previously analyzed samples, yielding results of 100% sensitivity and 90.48% specificity in detection expansions of CGG (>30) repeats in the gene. This method is appropriate for the quick determination of allelic changes in the gene, screening a population, and identifying mutations or premutation carriers in a population with intellectual disabilities of an unknown cause.
本研究的目的是验证用于确定该基因动态突变的直接三联体引物PCR方法（dTP-PCR），并比较dTP-PCR方法和Southern印迹分析的结果。通过直接三联体引物PCR方法和熔解曲线分析来确定该基因中CGG重复序列的数量。使用具有已知CGG重复序列数量的对照样品来确定CGG重复序列正常与排列之间的临界温度。根据DNA熔解曲线将所有患者分为四类。通过40个先前分析的样本确定了该检测方法的临床性能，在检测该基因中CGG（>30）重复序列的扩增时，灵敏度为100%，特异性为90.48%。该方法适用于快速确定该基因的等位基因变化、筛查人群以及在病因不明的智力残疾人群中识别突变或前突变携带者。
