论文部分内容阅读
目的研究江苏徐州地区肝豆状核变性(又称Wilson’病,WD)患者ATP基因12、14、18外显子突变情况,以期建立徐州地区WD的ATP基因突变热点的检测平台。方法采集45例WD患者及50例正常人外周血,提取基因组DNA,PCR扩增ATP7B第12、14、18号外显子,12号外显子扩增产物进行Tail内切酶酶切分析;然后对所有病人及正常人DNA外显子扩增产物进行直接测序,进而与临床表型做相关分析。结果 45例WD病人中,共发现6例突变。12号显子无1例存在Tail酶切反应异常,测序共发现2855G→A多态(即Arg952Lys)9例,其中1例合并12号外显子2828G→A杂合突变(即Gly943Asp),18号外显子发现3884C→T(即Ala1295Val)杂合突变5例,其中2例合并3889G→A多态(即Val1297lle),14号外显子未见突变。结论 18号外显子可能是徐州地区WD患者的基因突变热区,可作为徐州地区WD可疑患者的优先筛选区域之一。12、14号外显子可能不是徐州地区WD患者的基因突变热区。
Objective To study the mutation of exons 12, 14 and 18 of Wilson ’s disease (Wilson’ s disease, WD) in Xuzhou district of Jiangsu Province in order to establish a detection platform for hot spots of ATP gene mutations in WD in Xuzhou area. Methods Peripheral blood was collected from 45 patients with WD and 50 normal controls. The genomic DNA was extracted and the exon 12, exon 18, exon 12 and exon 12 of PCR-amplified products were analyzed by Tail endonuclease digestion. All patients and normal DNA exon amplification products were directly sequenced, and then do with the clinical phenotype correlation analysis. Results 45 cases of WD patients, found a total of 6 mutations. There was no abnormality of Tail digestion reaction in 12 cases. There were 9 cases of 2855G → A polymorphism (Arg952Lys), including 1 case of 2828G → A heterozygous mutation of exon 12 (Gly943Asp) There were 5 heterozygous mutations of 3884C → T (Ala1295Val) in exon, of which 2 cases had 3889G → A polymorphism (Val1297lle) and no mutation in exon 14. Conclusion Exon 18 may be a hot mutation in WD patients in Xuzhou area, which may be one of the preferential screening regions for WD suspicious patients in Xuzhou area. Exons 12 and 14 may not be the hot mutation in WD patients in Xuzhou.