目的

探讨利用新的高分辨率熔解曲线分析法检测结直肠癌中KRAS基因突变的可行性。

方法

首先采用直接测序法检测15例III期结直肠癌患者中KRAS基因第2外显子上第12和13号密码子的突变情况，继而以高分辨率熔解曲线分析法对直接测序的结果进行验证。

结果

经直接测序，确认15例结直肠癌患者中有6例发生了KRAS基因突变（40%），突变患者的基因型为：GGT>GAT型占2例，GGT>GTT占2例，GGC>GAC型占1例，GGC>GAA占1例。通过高分辨率熔解曲线分析法对以上15例标本进行进一步验证，证实所有患者的突变情况与直接测序方法的结果完全吻合。

结论

新的高分辨率熔解曲线分析法在检测KRAS基因突变时比直接测序法具有耗时短，易操作的优点，并且检测成本低廉，适合在临床开展。

Objective

To evaluate the feasibility and sensitivity of detecting the mutations of KRAS in colorectal carcinoma patients with high resolution melting curve analysis.

Methods

Direct sequencing was applied to detect the mutations of KRAS codon 12 and 13 of 15 patients with stage III colorectal carcinoma.Then the results were confirmed with high resolution melting curve analysis.

Results

By direct sequencing, 6 of 15 patients(40%)were found to carry KRAS mutations,and the distributions were 2 patients harboring GGT>GAT mutation,2 patients harboring GGT>GTT mutation,1 patient harboring GGC>GAC mutation and 1 patient harboring GGC>GAA mutation.By high resolution melting curve analysis,consistent results of the patients with mutations or without mutations were got.

Conclusions

The new high resolution melting curve analysis is more efficient,more convenient than direct sequencing.Moreover,it is a low-cost test,which is suitable for clinic test.