摘要
目的对4个肾上腺脑白质营养不良(ALD)家系进行基因突变分析。方法应用变性 HPLC 技术检测了4个 ALD 家系的 ABCD1基因,并通过核苷酸序列分析确证突变位点。结果所有患儿的母亲,患儿2、患儿3、患儿4弟弟及其表弟与正常对照的 PCR 产物混合物,均出现变性HPLC 洗脱双峰,而正常对照均为单峰,提示相应基因片段存在突变位点。上述出现异源双峰的 PCR产物经直接测序,证实了相关突变的存在:患儿1母亲为 fs Glu 471突变携带者;患儿2存在 S108X 突变;患儿3存在 R617C 突变;患儿4弟弟和表弟均存在 A141T 突变。结论变性 HPLC 能有效地检测ABCD1基因突变,并为进一步地基因突变筛查与产前诊断提供依据。
Objective To detect the mutations of ABCD1 gene by denaturing high performance liquid chromatography （DHPLC）. Methods DHPLC was used to examine the ABCD1 gene in four Chinese families with X-liked adrenoleukodystrophy （X-ALD）. The suspectable mutations were comfirmed by DNA sequencing. Results In the abnormal DHPLC profiles, double peaks were found in PCR products of all patients＇ mothers, the mixed PCR products of corresponding normal controls and patient 2,3, brother or cousin of patient 4, which indicated the presence of sequence alterations in ABCD1 gene. All the alterations were confirmed by sequencing, which revealed four distinct mutations in ABCD1 genes of the four pedigrees. Thus, a mutation of fs Glu 471 was found in one ABCD1 allele of the mother of patient 1. The mutations of S108X and R617C were identified in ABCD1 gene from patient 2 and patient 3, respectively. While a mutation of A141T was detected in ABCD1 gene from the brother and cousin of patient 4. Conclusion The DHPLC protocol could act as an efficient assay for mutation detecting of X-ALD families, and its application would be beneficial to further mutation screening and prenatal diagnosis.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2007年第2期157-160,共4页
Chinese Journal of Laboratory Medicine
关键词
肾上腺脑白质营养不良
分子诊断技术
基因
色谱法
高压液相
Adrenoleukodystrophy
Molecular diagnostic techniques
Gene
Chromatography, high pressure liquid
