摘要
目的优选出与运动神经元生存(survival motor neuron,SMN)基因紧密连锁且多态信息含量丰富的短串联重复序列(STR)位点,并将其应用于脊髓性肌萎缩症(spinal muscular atrophy,SMA)产前基因诊断中。方法用 PCR 扩增与 SMN 基因紧密连锁的11个 STR 位点,聚丙烯酰胺凝胶电泳(PAGE)检测,银染法显色分析结果。通过多态信息含量(PIC)大小优选 STR 位点,并利用优选出的 STR 位点分别采用 PCR-PAGE 及基因扫描技术对6个 SMA 家系(包括父母、先证者和胎儿)进行连锁分析。结果我们从11个 STR 位点中优选出了3个位点(D5S435、D5F149、D5S351),这3个位点在100名健康人中分别检测出8、19、18种多态性片段,其 PIC 值分别为0.84、0.91、0.92。应用上述3个位点对6个 SMA 家系进行产前诊断连锁分析,在6名胎儿中发现4名携带者和2名健康者。结论通过优选出的3个 STR 位点可快速进行 SMA 产前基因诊断,准确地排除母血污染,而且可在胎儿中甄别出健康个体与基因携带者,进一步完善了 SMA 产前基因诊断体系。
Objective To optimize the short tandem repeats （STR） which link closely to survival motor neuron （SMN） and have redundant polymorphism information contents, and to use these STR in the prenatal diagnosis of spinal muscular atrophy （SMA）. Methods Eleven STR loci （ D5S435, D5FI53, D5F151, D5S637, D5S1413, D5S125, D5S464, D5S1556, D5F149, D5S351, MAP1B-5＇） were amplified by PCR. Then the PCR products were detected by polyacrylamide gel electrophoresis （PAGE） and analyzed by silver staining. STR loci were evaluated and optimized by their PIC values. PCR-PAGE and gene scan were combined to make genetic link analysis for SMA families based on the optimized STR. Results Three STR loci （D5S435, D5F149 and D5S351 ） were selected with 8, 19 and 18 polymorphic fragments detected respectively in 100 normal individuals. Their PIC values were 0. 84, 0. 91 and 0. 92 respectively. Four carriers and 2 normal individuals were detected from 6 SMA families with linkage analysis by using the 3 STR. Conclusion This genetic diagnosis system based on the 3 STR loci can provide rapid prenatal diagnosis for SMA families, can eliminate maternal blood contamination, and also can discriminate carriers from normal individuals in the fetuses, which makes the prenatal diagnosis system of SMA perfect.
出处
《中华神经科杂志》
CAS
CSCD
北大核心
2007年第7期460-464,共5页
Chinese Journal of Neurology
基金
国家自然科学基金(30670730)
福建省自然科学基金计划资助项目(2006J0086)
福建省卫生厅青年科研课题资助项目(2005-1-4)
福建医科大学青年教师科研基金(FJCXQ04007)
福建省重大科技基金资助项目(2002Y001)
关键词
肌萎缩
脊髓性
串联重复序列
产前诊断
电泳
聚丙烯酰氨凝胶
Muscular atrophy, spinal
Tandem repeat sequences
Prenatal diagnosis
Electrophoresis, polyacrylamide gel
