KRAS基因突变直接测序检测方法的建立

目的 建立一种适用于临床的KRAS基因直接测序突变检测方法.方法 以KRAS基因12、13密码子为突变检测靶位点设计特异性扩增、测序引物,以已知12、13密码子野生型、突变型样品分别做为阴、阳性对照品,建立KRAS基因突变直接测序检测方法,并行方法学评估.结果 成功建立了KRAS基因突变直接测序检测方法,该方法检测灵敏度达3.9 ng/μl,重复性良好.检测17例结直肠癌样品,突变率为23.5％.结论 本研究成功建立了可用于临床样品检测的KRAS基因突变直接测序检测方法.

Establishment of a sequencing method for detecting KRAS gene mutations

Objective To establish a sequencing method for detecting KRAS gene mutations. Methods A sequencing method for KRAS gene mutations detection was established with specific primers that targeted hotspot mutation region （codonl2 and codonl3）, and wild-type and mutation samples were selected as negative sample and positive sample respectively. Then the performance of the method was assessed. Results A sequencing method for KRAS gene mutations detection was established successfully which owned high sensitivity （3.9 ng/μl） and good repeatability. The mutation rate was 23.5% in 17 colorectal cancer samples. Conclusion The sequencing method can be used in clinical samples to detect KRAS genemutations.