焦磷酸测序法检测BRAF V600E突变
Detection of BRAF V600E mutation by pyrosequencing.
作者信息
Tan Yi Hui, Liu Yanqun, Eu Kong Weng, Ang Pei Woon, Li Wei Qi, Salto-Tellez Manuel, Iacopetta Barry, Soong Richie
机构信息
Oncology Research Institute, National University of Singapore, Singapore.
出版信息
Pathology. 2008 Apr;40(3):295-8. doi: 10.1080/00313020801911512.
INTRODUCTION
Detection of the V600E hotspot mutation in BRAF oncogene is extremely useful for the screening of hereditary non-polyposis colorectal cancer (Lynch's syndrome) and for the prediction of sensitivity to MEK inhibitors. Here we describe a method for detecting this mutation based upon pyrosequencing technology.
METHODS
The efficiency of pyrosequencing for detecting BRAF V600E mutations was compared with the conventional dideoxy sequencing method in 12 tumour cell lines and in 108 colorectal tumours.
RESULTS
The results from pyrosequencing were 100% concordant with those from dideoxy sequencing. This method was capable of detecting BRAF V600E mutations at a much lower ratio of mutant to wild-type alleles (1:50) than dideoxy sequencing (1:5) while being considerably faster and less expensive.
CONCLUSIONS
Pyrosequencing offers a specific, sensitive, rapid and cost-effective alternative to dideoxy sequencing for the detection of BRAF V600E mutations in clinical tumour specimens.
引言
检测BRAF癌基因中的V600E热点突变对于遗传性非息肉病性结直肠癌（林奇综合征）的筛查以及对MEK抑制剂敏感性的预测极为有用。在此，我们描述一种基于焦磷酸测序技术检测该突变的方法。
方法
在12个肿瘤细胞系和108个结直肠癌肿瘤中，将焦磷酸测序检测BRAF V600E突变的效率与传统双脱氧测序方法进行比较。
结果
焦磷酸测序结果与双脱氧测序结果100%一致。该方法能够以比双脱氧测序（1:5）低得多的突变型与野生型等位基因比例（1:50）检测BRAF V600E突变，同时速度更快且成本更低。
结论
对于临床肿瘤标本中BRAF V600E突变的检测，焦磷酸测序为双脱氧测序提供了一种特异、灵敏、快速且经济高效的替代方法。
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