Diagnosis of Fanconi Anemia: Mutation Analysis by Multiplex Ligation-Dependent Probe Amplification and PCR-Based Sanger Sequencing.
Diagnosis of Fanconi Anemia: Mutation Analysis by Multiplex Ligation-Dependent Probe Amplification and PCR-Based Sanger Sequencing.
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作者：
Gille JJ;Floor K;Kerkhoven L;Ameziane N;Joenje H;de Winter JP
Fanconi贫血(FA)是一种罕见的遗传性疾病，以发育缺陷、身材矮小、骨髓衰竭和恶性肿瘤的高风险为特征。FA是异质性的：到目前为止已经区分了15个遗传亚型。FA的临床诊断需要通过在染色体断裂试验中测试细胞对交联剂的敏感性来确认。作为第二步，DNA检测可以用来阐明患者的基因亚型，并识别家族性突变。这一知识可以进行植入前遗传诊断(PGD)，并能够在未来怀孕时进行产前DNA测试。虽然通过下一代测序同时测试所有FA基因在不久的将来将成为可能，但这项技术不会立即适用于所有实验室。此外，在创始人突变较强的人群中，使用桑格测序和MLPA进行有限测试将是一种成本效益高的替代方案。我们描述了筛查FANCA、FANCB、FANCC、FANCE、FANCF和FANCG的策略和优化条件，并介绍了自2008年以来在我们的诊断服务中转诊的54名患者的结果。此外，还讨论了家系遗传咨询和携带者筛查的后续工作。
Fanconi anemia (FA) is a rare inherited disease characterized by developmental defects, short stature, bone marrow failure, and a high risk of malignancies. FA is heterogeneous: 15 genetic subtypes have been distinguished so far. A clinical diagnosis of FA needs to be confirmed by testing cells for sensitivity to cross-linking agents in a chromosomal breakage test. As a second step, DNA testing can be employed to elucidate the genetic subtype of the patient and to identify the familial mutations. This knowledge allows preimplantation genetic diagnosis (PGD) and enables prenatal DNA testing in future pregnancies. Although simultaneous testing of all FA genes by next generation sequencing will be possible in the near future, this technique will not be available immediately for all laboratories. In addition, in populations with strong founder mutations, a limited test using Sanger sequencing and MLPA will be a cost-effective alternative. We describe a strategy and optimized conditions for the screening of FANCA, FANCB, FANCC, FANCE, FANCF, and FANCG and present the results obtained in a cohort of 54 patients referred to our diagnostic service since 2008. In addition, the follow up with respect to genetic counseling and carrier screening in the families is discussed.