摘要
目的 建立一种适用于临床样品检测的BRAF基因V600E突变的检测方法.方法 利用已知BRAF基因野生型、突变型样品,以BRAF基因15外显子V600E突变为靶位点设计特异性扩增引物和测序引物,建立BRAF基因V600E突变的SYRB Green PCR-Sanger测序检测方法,并对该方法进行方法学评价和21例结直肠癌临床样品检测.结果 成功建立了BRAF基因V600E突变SYRB GreenPCR-Sanger测序检测方法,该方法的灵敏度达5.0×101拷贝/ul,检测线性范围5×101～ 5×107拷贝/ul,且检测的重复性好.检测21例结直肠癌临床样品,突变率为9.5％（2/21）.结论 成功建立了BRAF基因V600E突变SYRB Green PCR-Sanger测序检测方法,该方法可用于临床样品的检测.
Objective To establish a clinically feasible method for detection of BRAF gene V600E mutation.Methods Based on the known sequences of BRAF wild-type and mutation samples,specific polymerase chain reaction and sequencing primers targeting at the BRAF gene 15 exon V600E mutation were designed.This allowed the establishment of SYRB Green PCR-Sanger sequencing technique for the detection of BRAF V600E mutation,for methodological evaluation and assay of 21 colorectal cancer samples.Results The SYRB Green PCR-Sanger sequencing method for detection of BRAF gene V600E mutation was established successfully,which harbored high assay sensitivity （5.0 × 101 copies/μl） and repeatability and acceptable linear range （5× 101 to 5× 107 copies/μl）.The mutation rate was 9.5％ （2/21） in 21 colorectal cancer samples.Condnsion The SYRB Green PCR-Sanger sequencing method for detection of BRAF gene V600E mutation is successfully established and can be applied in assessments for clinical samples.
出处
《中华生物医学工程杂志》
CAS
2014年第3期215-218,共4页
Chinese Journal of Biomedical Engineering
关键词
突变
聚合酶链反应
寡核苷酸序列分析
基因
BRAF
Mutation Polymerase chain reaction Oligonucleotide array sequence analysis Genes,BRAF
