Moghrabi Nabil N, Johnson Monique A, Yoshitomi Marvin J, Zhu Xiaoman, Al-Dhalimy Muhsen J, Olson Susan B, Grompe Markus, Richards C Sue
Department of Molecular and Medical Genetics, Clinical Genetics Laboratories, Oregon Health & Science University, 3181 S.W. Sam Jackson Park Road, Portland, OR 97239, USA.
Genet Med. 2009 Mar;11(3):183-92. doi: 10.1097/GIM.0b013e318193ba67.
Fanconi anemia is a genetically heterogeneous chromosomal breakage disorder exhibiting a high degree of clinical variability. Clinical diagnoses are confirmed by testing patient cells for increased sensitivity to crosslinking agents. Fanconi anemia complementation group assignment, essential for efficient molecular diagnosis of the disease, had not been validated for clinical application before this study. The purpose of this study was (1) confirmation of the accuracy of Fanconi anemia complementation group assignment to Group A (FANCA) and (2) development of a rapid mutation detection strategy that ensures the efficient capture of all FANCA mutations.
Using fibroblasts from 29 patients, diagnosis of Fanconi anemia and assignment to complementation Group A was made through breakage analysis studies. FANCA coding and flanking sequences were analyzed using denaturing high pressure liquid chromatography, sequencing, and multiplex ligation-dependent probe amplification. Patients in which two mutations were not identified were analyzed by cDNA sequencing. Patients with no mutations were sequenced for mutations in FANCC, G, E, and F.
Of the 56 putative mutant alleles studied, 89% had an identifiable FANCA pathogenic mutation. Eight unique novel mutations were identified.
Complementation assignment to Group A was validated in a clinical laboratory setting using our FANCA rapid molecular testing strategy.
范可尼贫血是一种具有遗传异质性的染色体断裂障碍疾病，临床变异性高。通过检测患者细胞对交联剂的敏感性增加来确诊临床诊断。范可尼贫血互补组的分类对于该疾病的有效分子诊断至关重要，在本研究之前尚未在临床应用中得到验证。本研究的目的是（1）确认范可尼贫血互补组分类为A组（FANCA）的准确性，以及（2）开发一种快速突变检测策略，以确保有效捕获所有FANCA突变。
使用29例患者的成纤维细胞，通过断裂分析研究进行范可尼贫血的诊断和互补组A的分类。使用变性高压液相色谱、测序和多重连接依赖探针扩增分析FANCA编码和侧翼序列。未鉴定出两个突变的患者通过cDNA测序进行分析。未发现突变的患者对FANCC、G、E和F的突变进行测序。
在所研究的56个推定突变等位基因中，89%有可识别的FANCA致病突变。鉴定出8个独特的新突变。
使用我们的FANCA快速分子检测策略，在临床实验室环境中验证了互补组分类为A组。
