Soltysova Andrea, Tothova Tarova Eva, Ficek Andrej, Baldovic Marian, Polakova Helena, Kayserova Hana, Kadasi Ludevit
Department of Molecular Biology, Faculty of Natural Sciences, Comenius University, Mlynska dolina Bratislava 842 15, Slovakia.
Institute for Clinical and Translational Research, Biomedical Research Center, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava 845 05, Slovakia.
Clin Respir J. 2018 Mar;12(3):1197-1206. doi: 10.1111/crj.12651. Epub 2017 Jun 5.
Cystic fibrosis (CF) has one of the longest histories in hereditary disease molecular diagnostics. However, identification of causative mutations in the CFTR gene is complicated by over 2000 currently identified mutations; with more still being discovered. Knowledge of mutation spectrum may improve effective routine diagnostics and is obligatory in mutation-specific treatment.
This study presents comprehensive mutation screening of the CFTR gene; with 275 unrelated, clinically confirmed and treated cystic fibrosis (CF) patients diagnosed in 25 years genetic testing in Slovakia.
Detection of the most common CFTR mutations was performed by ELUCIGENE 29 and ELUCIGENE CF EU2 kits. HRM and dHPLC mutation screening methods with subsequent Sanger sequencing were applied for minor mutation screening, and MLPA analysis for deletion/duplication detection.
A total of 70 different mutations were identified, from which the most common mutation F508del accounted for 60.36% of all disease alleles and 8 mutations have currently been observed only in Slovak patients. Two large deletions identified on chromosomes 2 and 22 were further characterized to identify breakpoints. Based on mutation screening results and neonatal screening we estimated incidence in Slovakian newborns at approximately 1:6000-7000.
In our study, we identified mutations in 98.54% of all disease chromosomes, while 86.54% were identified using ELUCIGENE kits, 0.54% by MLPA analysis and 11.46% by sequencing analysis. Knowledge of the mutation spectrum in genetically diagnosed patients improves possibilities of genetic counseling and cascade screening in the affected families and Slovak population.
囊性纤维化（CF）在遗传性疾病分子诊断领域拥有悠久的历史。然而，CFTR基因中致病突变的鉴定因目前已发现的2000多种突变而变得复杂，且仍有更多突变不断被发现。了解突变谱可能会改善有效的常规诊断，并且在针对特定突变的治疗中是必不可少的。
本研究对CFTR基因进行了全面的突变筛查，涉及斯洛伐克25年基因检测中确诊并接受治疗的275例无亲缘关系的临床确诊囊性纤维化（CF）患者。
使用ELUCIGENE 29和ELUCIGENE CF EU2试剂盒检测最常见的CFTR突变。采用高分辨率熔解曲线分析（HRM）和变性高效液相色谱（dHPLC）突变筛查方法及随后的桑格测序进行罕见突变筛查，并采用多重连接探针扩增（MLPA）分析检测缺失/重复。
共鉴定出70种不同的突变，其中最常见的突变F508del占所有致病等位基因的60.36%，目前已观察到8种突变仅在斯洛伐克患者中出现。对在2号和22号染色体上鉴定出的两个大片段缺失进行了进一步表征以确定断点。根据突变筛查结果和新生儿筛查，我们估计斯洛伐克新生儿的发病率约为1:6000 - 7000。
在我们的研究中，我们在所有致病染色体中鉴定出98.54%的突变，其中86.54%通过ELUCIGENE试剂盒鉴定，0.54%通过MLPA分析鉴定，11.46%通过测序分析鉴定。了解基因诊断患者的突变谱可提高在受影响家庭和斯洛伐克人群中进行遗传咨询和级联筛查的可能性。
