Institute of Clinical Research, University of Southern Denmark, Odense C, Denmark.
J Hum Genet. 2012 Jul;57(7):453-8. doi: 10.1038/jhg.2012.56. Epub 2012 Jun 14.
This study aimed to identify the underlying genetic mutation in patients with hypophosphatemic rickets (HR). Genomic DNA was analysed for mutations in PHEX, FGF23 and CLCN5 by polymerase chain reaction (PCR) followed by denaturing high-performance liquid chromatography (dHPLC). Bi-directional sequencing was performed in samples with deviating chromatographic profiles. DMP1 and SLC34A3 were sequenced, only. In addition, a multiplex ligation-dependent probe amplification (MLPA) analysis was performed to detect larger deletions/duplications in PHEX or FGF23. Familial cases accounted for 12 probands while 12 cases were sporadic. In 20 probands, mutations were detected in PHEX of which 12 were novel, and one novel frameshift mutation was found in DMP1. Three PHEX mutations were identified by the MLPA analysis only; that is, two large deletions and one duplication. No mutations were identified in FGF23, SLC34A3 or CLCN5. By the methods used, a disease causing mutation was identified in 83% of the familial and 92% of the sporadic cases, thereby in 88% of the tested probands. Genetic analysis performed in HR patients by PCR, dHPLC, sequencing and in addition by MLPA analysis revealed a high identification rate of gene mutations causing HR, including 12 novel PHEX and one novel DMP1 mutation.
本研究旨在鉴定低血磷性佝偻病（HR）患者的潜在遗传突变。通过聚合酶链反应（PCR）后变性高效液相色谱（dHPLC）分析 PHEX、FGF23 和 CLCN5 的基因突变。在具有偏离色谱图谱的样本中进行双向测序。仅对 DMP1 和 SLC34A3 进行测序。此外，还进行了多重连接依赖性探针扩增（MLPA）分析，以检测 PHEX 或 FGF23 中的较大缺失/重复。家族病例占 12 个先证者，而散发病例为 12 例。在 20 个先证者中，检测到 PHEX 突变，其中 12 个为新突变，DMP1 中发现一个新的移码突变。仅通过 MLPA 分析鉴定出 3 个 PHEX 突变，即两个大片段缺失和一个重复。未在 FGF23、SLC34A3 或 CLCN5 中发现突变。通过使用的方法，在家族性和散发性病例中分别有 83%和 92%的病例确定了致病突变，因此在 88%的检测先证者中确定了致病突变。通过 PCR、dHPLC、测序以及 MLPA 分析对 HR 患者进行的遗传分析显示，导致 HR 的基因突变的识别率很高，包括 12 个新的 PHEX 和一个新的 DMP1 突变。
