论文部分内容阅读
目的对中国人Ⅰ型神经纤维瘤病(NF1)基因20、28、29、39号外显子进行基因突变分析及评价聚合酶链反应-单链构象多态/异源双链(PCR-SSCP/HA)技术在NF1基因诊断中的价值。方法应用PCR-SSCP/HA技术,结合DNA测序,筛查56例患者NF1基因20、28、29、39号外显子的突变或多态性。结果在20号外显子发现一个家系父子3人的SSCP/HA出现泳动异常,DNA测序证实为20号外显子上T→G的杂合突变即Leul141Arg错义突变:发现1例患者28号外显子的SSCP/HA检测有泳动异常,测序证实为28号外显子5’端上游的第28位核苷酸G→T杂合;29、39号外显子未检测到泳动异常的条带。结论联合应用SSCP/HA的方法可提高基因突变检测敏感度及检出率,NF1基因20、28、29、39号外显子不是突变热点或突变率相对较高的区域。
Objective To analyze the gene mutations of exon 20, 28, 29 and 39 of Chinese type Ⅰ neurofibromatosis (NF1) gene and evaluate the effect of PCR-SSCP / HA) technology in the diagnosis of NF1 gene value. Methods The mutations or polymorphisms of exon 20, 28, 29 and 39 of NF1 gene in 56 patients were screened by PCR-SSCP / HA combined with DNA sequencing. Results In exon 20, SSCP / HA of 3 pedigrees was found to be abnormal in motility. DNA sequencing confirmed the heterozygous mutation of T → G on exon 20, ie, the mutation of Leul141Arg. One patient, The sub-SSCP / HA test showed abnormality of migration. Sequencing confirmed that the 28th nucleotide G → T was upstream of the 5 ’end of exon 28 and no abnormal migration was detected in exons 29 and 39 . Conclusion The combined application of SSCP / HA method can improve the sensitivity and detection rate of gene mutation detection. Exon 20, 28, 29 and 39 of NF1 gene are not hot spots or mutation areas with relatively high mutation rate.