title: A novel method for detection of mutation in epidermal growth factor receptor.
abstract: 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.
  
  
  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 ΔCt 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 specificity.
  
  
  CONCLUSIONS
  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.
authors: Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China.
authors: Jinyin
authors: J
authors: Zhao
authors: Jing
authors: J
authors: Zhao
authors: Jie
authors: J
authors: Huang
authors: Yan
authors: Y
authors: Chen
authors: Jun
authors: J
authors: Jiang
authors: Weili
authors: W
authors: Wu
authors: Pengzhi
authors: P
authors: Wang
authors: Licheng
authors: L
authors: Liu
authors: Longyun
authors: L
authors: Li
authors: Lin
authors: L
authors: Wu
authors: Mengzhao
authors: M
authors: Wang
authors: Weijun
authors: W
authors: Chen
mesh_terms: D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D004252:DNA Mutational Analysis / Q000379:methods; D017931:DNA Primers / Q000138:chemical synthesis; D066246:ErbB Receptors / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D066246:ErbB Receptors