腓骨肌萎缩症1A型基因重复的检测

Detection of duplication in Charcot-Marie-Tooth disease type 1A

目的 探索适合临床应用的检测腓骨肌萎缩症 1A型 (CMT1A)基因重复的有效方法。方法 联合应用多聚酶链反应 (PCR)加双酶切方法和短串联重复序列 (STR)分析检测CMT病人 17p11.2～ 12上的基因重复。共检测 30例无亲缘关系的CMT1病人和 10例无亲缘关系的CMT2病人及40例对照。结果 PCR 双酶切法的基因重复检出率为 46 .7% (14 30 ) ;STR分析的基因重复检出率为5 3.3% (16 30 ) ;联合应用上述两种方法的基因重复检出率为 70 .0 % (2 1 30 )。 10例无亲缘关系的CMT2病人和 40例对照组中 ,两种方法均未发现重复。结论 PCR 双酶切法检测CMT1A基因重复敏感、快速、准确 ,适于临床推广应用 ;联合应用PCR 双酶切 +STR分析 ,可提高CMT1A基因重复检出率。

Objective To study the routine methods that can be easily used in clinics to detect the Charcot Marie Tooth (CMT) disease gene duplication. Method Polymerase chain reaction(PCR) combined with restriction enzyme digestion and amplification of short tandem repeat (STR) sequence were used to detect gene duplication on chromosome 17p11.2～12 in 30 CMT1 patients and 10 CMT2 patients coming from unrelated families. 40 controls were also detected. Results 46.7%(14/30) of CMT1 patients were identified to have specific junction fragments. 53.3%(16/30)of them were identified as duplication by STR analysis. 70.0%(21/30) of CMT1 patients were identified to have gene duplication using both methods. Duplication was not identified in 10 unrelated CMT2 patients and 40 controls. Conclusion The PCR combined with restriction enzyme digestion represented a relatively sensitive and accurate method for detecting gene duplication in CMT1A cases for clinical diagnosis. The detecting rate of duplication can be increased using both restriction enzyme digestion of PCR products and STR methods.