CAJ | 학술논문

目的通过对1例疑似肾上腺脑白质营养不良(ALD)患者及该家系其他成员进行ALD基因分析来明确诊断. 方法从患者和家庭成员的外周血白细胞提取总RNA和基因组DNA:应用RT-PCR技术,首先对先证者cDNA的ABCD1基因编码区进行序列分析,寻找突变位点,再通过PCR和直接测序等方法进一步确证该突变位点;同时对该家系其他成员的相应基因组DNA进行ABCD1基因分析. 结果先证者的ABCD1基因第656(G)、657(A)位碱基缺失,造成移码突变fs R89,可以确诊为ALD.该家系其他成员基因突变分析结果为:先证者表弟存在与先证者相同的突变,为ALD半合子;先证者的母亲、小姨、表妹均为ALD携带者;先证者姐姐为ABCD1正常基因型. 结论在中国人ALD患者中发现了1个新的ABCD1基因突变(fs R89),ABCD1基因突变分析是诊断X-ALD最可靠的方法 .
목적 통과대1례의사신상선뇌백질영양불량(ALD)환자급해가계기타성원진행ALD기인분석래명학진단. 방법 종환자화가정성원적외주혈백세포제취총RNA화기인조DNA:응용RT-PCR기술,수선대선증자cDNA적ABCD1기인편마구진행서렬분석,심조돌변위점,재통과PCR화직접측서등방법 진일보학증해돌변위점;동시대해가계기타성원적상응기인조DNA진행ABCD1기인분석. 결과 선증자적ABCD1기인제656(G)、657(A)위감기결실,조성이마돌변fs R89,가이학진위ALD.해가계기타성원기인돌변분석결과 위:선증자표제존재여선증자상동적돌변,위ALD반합자;선증자적모친、소이、표매균위ALD휴대자;선증자저저위ABCD1정상기인형. 결론 재중국인ALD환자중발현료1개신적ABCD1기인돌변(fs R89),ABCD1기인돌변분석시진단X-ALD최가고적방법 .
Objective To identify the ABCD1 gene mutation in a patient suspected with adrenoleukodystrophy (ALD) and perform its gene analysis in his family members to make a definite diagnosis. Methods Total RNA and genomic DNA were extracted from the leukocytes of peripheral blood in the proband and the family members. The ABCD1 coding region of cDNA in the proband was amplified and sequenced. Mutational site in the ABCD1 gene of the proband was further confirmed by PGR and direct sequencing; at the same time, the mutation in the ABCD1 gene of the genomic DNA in the family members was analyzed by direct sequencing. Results Two bases deletions (656_657delGA)were identified and the corresponding mutation of fs R89 was detected in the ABCD1 gene of the proband, which could make the definite diagnosis of ALD that belonged to adrenomyeloneuropathy. The same gene mutation (ALD hemizygote) was noted in his cousin; his mother, younger sister of his mother and his younger female cousin were noted as the ALD carrers. His older sister was noted as ABCD1 normal genotype. Conclusions A novel ABCD1 gene mutation (fs R89) was identified in Chinese patient with ALD. Molecular testing is an effective way in making diagnosis on patient suspected as having X-linked ALD.