高分辨熔解曲线技术鉴定多药耐药基因外显子单核苷酸多态性  

High resolution melting analysis for single nucleotide polymorphism of MDR1 gene exon

作　　者：徐伟[1] 夏之柏[2] 朱斌[1] 

机构地区：[1]苏州大学基础医学与生物科学学院,苏州215123 [2]广州中山大学附属第一医院神经外科,广州510080

出　　处：《中国优生与遗传杂志》2012年第2期15-16,71,共3页Chinese Journal of Birth Health & Heredity

摘　　要：目的高分辨熔解曲线技术(HRM)检测多药耐药基因(MDR1)外显子12单核苷酸多态性(SNP)。方法采用高分辨熔解技术对MDR1基因外显子12的SNP C1236T位点进行基因分型,以其-401C>T位点为例设计PCR扩增引物,按PCR扩增效率和熔解曲线进行退火温度、升温速度等条件的优化,并用此优化体系基因分型20例外周血标本,以测序验证。结果 20例标本经测序与检测结果一致。结论高分辨熔解曲线技术检测SNP是一种低成本、简便易行、常规化,高通量的基因分型方法,能用于大规模临床筛查。Objective： To study the practicality of using high resolution melting analysis for single nucleotide polymorphism （SNP） of MDR1 gene exonl2. Methods： SNP of MDR1 gene exonl2 was genotyped with HRM, and the PCR primers was designed on the SNP site -401C 〉 T of MDR1 gene exon. The PER amplification system was optimized, including the concentration of the amplification efficiency and production specificity of PCR. Twenty clinical samples from out - patients were genotyped using the optimized PCR system and the genotypes were validated with the sequencing. Results ： The 20 cases samples by sequencing and testing of HRM results are consistent. Conclusions ： The HRM technique for genotyping on SNP is a simple, economic, routine method and high - throughput that deserves to be spread in large- scale clinical screening.

关 键 词：高分辨熔解曲线 多药耐药基因 SNP 

分 类 号：R440[医药卫生—诊断学]