A novel method for detection of mutation in epidermal growth factor receptor

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• 作者：Jinyin  ; Zhao^a ;   ; ¹ ; Jing  ; Zhao^b ;   ; ¹ ; Jie  ; Huang^c ;   ; ¹ ; Yan  ; Chen^a ; Jun  ; Jiang^a ; Weili  ; Wu^a ; Pengzhi  ; Wang^d ; Licheng  ; Liu^d ; Longyun  ; Li^b ; Lin  ; Wu^a ; Mengzhao  ; Wang^b ;   ; ^{mengzhaowang@sina.com} ; Weijun  ; Chen^a ;   ; ^d ;   ; ^e ;   ; ^{chenwj@genomics.org.cn}
• 关键词：Mutation-enriched PCR ; ARMS TaqMan real-time PCR ; EGFR
• 刊名：Lung Cancer
• 出版年：2011
• 出版时间：November 2011
• 年：2011
• 卷：74
• 期：2
• 页码：226-232
• 全文大小：1423 K

文摘

Background

For the rapid and sensitive screening of epidermal growth factor receptor (EGFR) hot-spot mutations, we developed a novel method combining mutant-enriched PCR with amplification refractory mutation system (ARMS) TaqMan real-time PCR in a one-step reaction tube.

Methods and results

We designed two pairs of primers to enrich and genotyping each mutation (E746_A750del and L858R): nest primers and ARMS primers. Before the PCR assays were carried out, the restriction enzymes were used to cut wild alleles. The results showed that this method could detect mutant alleles mixed samples containing 0.1 % with a cutoff ¦¤Ct value of 12. We used this method in a survey of 73 non-small cell lung cancer (NSCLC) samples, detecting 14 mutant samples of E746_A750del and 12 mutant samples of L858R. The results well agreed with the results of DxS. All unmatched samples were identified by sequencing and the results showed that our method has high specificty.

Conclusion

The mutant-enriched ARMS TaqMan PCR could be useful in the detection of mutation in clinical samples containing only a small number of mutant alleles.