Rapid diagnosis of Miller-Dieker syndrome and isolated lissencephaly sequence by the polymerase chain reaction.
Rapid diagnosis of Miller-Dieker syndrome and isolated lissencephaly sequence by the polymerase chain reaction.
复制标题
通过聚合酶链式反应快速诊断 Miller-Dieker 综合征和孤立性无脑畸形序列。
DOI：
10.1007/bf00194237
复制
发表时间：
1990
期刊：
影响因子：
5.3
通讯作者：
Ledbetter,DH
中科院分区：
文献类型：
--
作者：
Batanian,JR;Ledbetter,SA;Wolff,RK;Nakamura,Y;White,R;Dobyns,WB;Ledbetter,DH
探针YNZ 22（D17 S5）是一种高度多态性的可变数目串联重复序列（VNTR）标记物，先前显示在所有Miller-Dieker综合征（MDS）患者中缺失，但在孤立性无脑序列（ILS）患者中未缺失。引物构建到YNZ 22的多态性重复区域侧翼的独特序列，用于通过聚合酶链式反应（PCR）扩增。118个正常个体的分析显示12个等位基因（70 bp重复单位的拷贝数不同），大小从168到938 bp不等。回顾性研究8例MDS和6例ILS患者与Southern印迹分析在所有情况下，除了一个一致。在后者中，患者及其母亲中的一个非常大的等位基因（重复单位的12个拷贝）在最初的尝试中未能扩增，但通过降低反应中使用的基因组DNA的浓度而成功扩增。对2例MDS和5例ILS患者成功进行了前瞻性研究，并通过Southern印迹分析证实了所有病例。从总样本中，限制性片段长度多态性（RFLP）分析是完全翔实的10例MDS患者中的4例，并显示在所有4例缺失。11例ILS患者中有9例是杂合子，因此YNZ 22未缺失。在Miller-Dieker区域开发额外多态性标记的引物将导致所有MDS和ILS患者的快速基于PCR的诊断方法。YNZ 22的PCR分型也将促进该标记在其他应用中的使用，包括遗传连锁、亲子鉴定和法医学研究以及肿瘤中杂合性缺失的分析。
Probe YNZ22 (D17S5) is a highly polymorphic, variable number tandem repeat (VNTR) marker previously shown to be deleted in all patients with the Miller-Dieker syndrome (MDS) but not in patients with isolated lissencephaly sequence (ILS). Primers were constructed to the unique sequence flanking the polymorphic, repetitive region of YNZ22 for amplification by the polymerase chain reaction (PCR). Analysis of 118 normal individuals revealed 12 alleles (differing in copy number of a 70-bp repeat unit) ranging in size from 168 to 938 bp. A retrospective study of eight MDS and six ILS patients was consistent with Southern blot analysis in all cases except one. In the latter, a very large allele (12 copies of the repeat unit) in a patient and her mother failed to amplify on initial attempts, but was successfully amplified by reducing the concentration of genomic DNA used in the reaction. Prospective studies on two MDS and five ILS patients were successfully performed and confirmed in all cases by Southern blot analysis. From the total sample, restriction fragment length polymorphism (RFLP) analysis was fully informative in four of ten MDS patients and showed a deletion in all four cases. Nine of eleven ILS patients were heterozygous and therefore not deleted for YNZ22. Development of primers for additional polymorphic markers in the Miller-Dieker region will lead to a rapid PCR-based diagnostic approach for all MDS and ILS patients. PCR typing of YNZ22 will also facilitate use of this marker in other applications, including genetic linkage, paternity and forensic studies, and analysis of loss of heterozygosity in tumors.