MLPA在常染色体显性遗传多囊肾病基因诊断中的应用

Application of MLPA in gene diagnosis of autosomal dominant polycystic kidney disease

目的探讨多重连接探针扩增技术(MLPA)在常染色体显性遗传多囊肾病(ADPKD)基因诊断中的应用。方法采用MLPA对20例ADPKD患者的PKD1基因和PKD2基因进行检测。MLPA检测结果显示单一外显子扩增或疑似扩增者采用RT-PCR检测验证;MLPA检测结果显示单一外显子缺失或疑似缺失者采用PCR检测验证,存在扩增产物者进行测序验证。结果 MLPA检测显示:1例患者单一外显子缺失(PKD1 Exon40),5例单一外显子疑似缺失(PKD1 Exon1、PKD1 Exon25、PKD2 Exon8、PKD2 Exon8、PKD1 Exon25),3例单一外显子疑似扩增(PKD1 Exon6、PKD1 Exon7、PKD1 Exon7)。经RT-PCR检测验证,1例患者单一外显子扩增(PKD1 Exon6);经PCR检测与测序验证,1例患者单一外显子错义突变(PKD1 Exon40),1例单一外显子缺失(PKD2 Exon8)。结论 MLPA为ADPKD的基因诊断提供了一种新的方法。

Objective To investigate the application of multiplex ligation-dependent probe amplification(MLPA) in the gene diagnosis of autosomal dominant polycystic kidney disease(ADPKD). Methods MLPA was employed to detect the PKD1 gene and PKD2 gene in 20 patients with ADPKD.Verification with RT-PCR was performed for those with single exon duplication or suspected duplication detected by MLPA.Those with single exon deletion or suspected deletion detected by MLPA were verified with PCR,and sequencing analysis was conducted in those with amplification products. Results One patient with single exon deletion(PKD1 Exon40),5 patients with single exon suspected deletion(PKD1 Exon1,PKD1 Exon25,PKD2 Exon8,PKD2 Exon8 and PKD1 Exon25) and 3 patients with single exon suspected duplication(PKD1 Exon6,PKD1 Exon7 and PKD1 Exon7) were detected by MLPA.One patient with single exon duplication(PKD1 Exon6) was verified by RT-PCR,and one patient with single exon missense mutation(PKD1 Exon40) and one patient with single exon deletion(PKD2 Exon8) were verified by PCR and sequencing analysis. Conclusion MLPA may serve as a new method for gene diagnosis of ADPKD.