Molecular analysis and prenatal prediction of spinal muscular atrophy in Chinese patients by the combination of restriction fragment length polymorphism analysis, denaturing high-performance liquid chromatography, and linkage analysis
Molecular analysis and prenatal prediction of spinal muscular atrophy in Chinese patients by the combination of restriction fragment length polymorphism analysis, denaturing high-performance liquid chromatography, and linkage analysis
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结合限制性片段长度多态性分析、变性高效液相色谱和连锁分析对中国患者脊髓性肌萎缩症进行分子分析和产前预测
DOI：
10.1001/archneur.64.2.225
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发表时间：
2007-02-01
影响因子：
--
通讯作者：
Wang, Ning
中科院分区：
文献类型：
--
作者：
Chen, Wan-Jin;Wu, Zhi-Ying;Wang, Ning
背景：脊髓性肌萎缩症（SMA）的难治性突出了SMA家族产前诊断的重要性。然而，目前应用于产前筛查的系统还很不完善。目的：优化分子检测方法，建立相对完善的产前筛查系统。设计、地点和患者：对来自77个SMA家庭的87名患者和132名父母进行了SMN 1突变筛查。对10个SMA家系的11例胎儿进行产前预测。所有样本均来自福建医科大学附属第一医院神经内科。主要指标：采用限制性片段长度多态性分析和变性高效液相色谱法（DHPLC）对87例患者及其父母进行SMN1缺失筛查。对无SMN1缺失的患者，采用实时荧光定量聚合酶链反应检测SMN1拷贝数，直接测序法筛查SMN的微小突变。应用限制性片段长度多态性分析、DHPLC和连锁分析对11例胎儿进行产前预测。采用DHPLC和实时荧光定量聚合酶链反应检测14例SMN1缺失的胎儿和14例父母的SMN1拷贝数和SMA携带者。流产胎儿和出生婴儿的结果再次证实了限制性片段长度多态性分析和DHPLC。结果：77例患者中有72例SMN1基因缺失，缺失率为93.5%。其他5个家族均未检测到细微突变。4只胎仔存在SMN1缺失并流产。其余7例胎儿，4例携带者和3例正常人，均在医生建议下出生。14名父母是携带者。流产胎儿和出生婴儿的再确认结果与产前预测完全一致。7出生的婴儿进行了随访，直到最近，所有的normal.Conclusions：限制性片段长度多态性分析，DHPLC和连锁分析的基础上的分子诊断系统是一种高效，准确的方法，非常适合于临床实验室的常规使用。
Background: The difficulties and incurability of spinal muscular atrophy (SMA) highlight the importance of prenatal diagnosis in families with SMA. However, the system applied in prenatal screening is far from perfect.Objectives: To optimize the molecular assays and establish a relatively perfect system for prenatal screening.Design, Setting, and Patients: A total of 87 patients and 132 parents from 77 families with SMA were screened for SMN1 mutations. Prenatal prediction was performed for 11 fetuses from 10 families with SMA. All of the samples to be tested were from the Department of Neurology, First Affiliated Hospital, Fujian Medical University, Fuzhou, China.Main Outcome Measures: All of the 87 patients and their parents were screened for SMN1 deletion by restriction fragment length polymorphism analysis and denaturing high-performance liquid chromatography (DHPLC). For those patients without the SMN1 deletion, the SMN1 copy numbers were detected by real-time fluorescence quantitative polymerase chain reaction and the subtle mutations of SMN were screened by direct sequencing. Prenatal prediction was performed by restriction fragment length polymorphism analysis, DHPLC, and linkage analysis for 11 fetuses. Furthermore, the SMN1 copy numbers and detected carriers of SMA were found by DHPLC and real-time fluorescence quantitative polymerase chain reaction in 14 parents and the fetuses without the SMN1 deletion. Results in aborted fetuses and born babies were reconfirmed by restriction fragment length polymorphism analysis and DHPLC. The born babies were followed up and physically examined twice a year.Results: The frequency of the SMN1 deletion we detected was 93.5% (72 of 77 patients). No subtle mutations were detected in the other 5 families. Four fetuses had the SMN1 deletion and were aborted. The other 7 fetuses, 4 carriers and 3 normal individuals, were born under suggestion by the physician. Fourteen parents were carriers. The reconfirmation of results in the aborted fetuses and born babies was completely consistent with prenatal prediction. The 7 born babies were followed up until recently and all were normal.Conclusions: The molecular diagnosis system based on restriction fragment length polymorphism analysis, DHPLC, and linkage analysis is an efficient and accurate method that is well suited for routine use in clinical laboratories.