摘要
目的应用扩增阻滞突变系统(amplification refractory mutationsystem,ARMS)排除ABCD1假基因的干扰,对肾上腺脑白质营养不良(adrenoleukodystrophy, ALD)家系进行基因突变分析。方法按ARMS引物设计原则设计上游引物(正常引物、突变引物)和下游公共引物,对家系成员的基因组DNA分别进行PCR扩增。结果R617C突变患者及其母亲的突变引物均扩增出特异性条带(107 bp) ,患者父亲和对照则未见条带,在基因组DNA水平证实了R617C突变的存在。结论双侧ARMS方法可以快速、有效地排除假基因干扰。
Objective To avoid the interference of ABCD1 pseudogenes, the amplification refractory mutation system （ARMS） was used to analyze the mutation of ABCD1 gene in the molecular diagnosis of X-linked adrenoleukedystrophy （ALD）. Methods The upstream primers （wild primer and mutation primer） were designed according to the principle of primer-design in ARMS. In addition, a common downstream primer was designed in the same way to discriminate ABCD1 gene from its prologous pseudogenes. The genomic DNA isolated from the peripheral blood leukocytes of the farnily members and normal controls was amplified by PCR. Results In double ARMS, a specific product of 107bp could be amplified from genomic DNA of the patient with R617C mutation in ABCD1 gene and his mother, while the same product was not found when the genomic DNA of the patient＇ s father and normal controls was used. Thus, the interference of ABCD1 pseudogenes in molecular diagnosis of ALD was excluded successfully at genomic DNA level. Conclusion Double ARMS is a quick and effective method to eliminate the interference of the pseudogenes in detecting ABCD1 gene mutations.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
2006年第5期558-560,共3页
Chinese Journal of Medical Genetics
