title: Quantum dots immunofluorescence histochemical detection of EGFR gene mutations in the non-small cell lung cancers using mutation-specific antibodies.
abstract: BACKGROUND
  Epidermal growth factor receptor (EGFR) mutation status plays an important role in therapeutic decision making for non-small cell lung cancer (NSCLC) patients. Since EGFR mutation-specific antibodies (E746-A750del and L858R) have been developed, EGFR mutation detection by immunohistochemistry (IHC) is a suitable screening test. On this basis, we want to establish a new screening test, quantum dots immunofluorescence histochemistry (QDs-IHC), to assess EGFR gene mutation in NSCLC tissues, and we compared it to traditional IHC and amplification refractory mutation system (ARMS).
  
  
  METHODS
  EGFR gene mutations were detected by QDs-IHC, IHC, and ADx-ARMS in 65 cases of NSCLC composed of 55 formalin-fixed, paraffin-embedded specimens and ten pleural effusion cell blocks, including 13 squamous cell carcinomas, two adenosquamous carcinomas, and 50 adenocarcinomas.
  
  
  RESULTS
  Positive rates of EGFR gene mutations detected by QDs-IHC, IHC, and ADx-ARMS were 40.0%, 36.9%, and 46.2%, respectively, in 65 cases of NSCLC patients. The sensitivity of QDs-IHC when detecting EGFR mutations, as compared to ADx-ARMS, was 86.7% (26/30); the specificity for both antibodies was 100.0% (26/26). IHC sensitivity was 80.0% (24/30) and the specificity was 92.31% (24/26). When detecting EGFR mutations, QDs-IHC and ADx-ARMS had perfect consistency (κ  =0.882; P<0.01). Excellent agreement was observed between IHC and ADx-ARMS when detecting EGFR mutations (κ  =0.826; P<0.01).
  
  
  CONCLUSIONS
  QDs-IHC is a simple and standardized method to detect EGFR mutations with its high sensitivity and specificity, as compared with real-time polymerase chain reaction. In addition, the development of specific antibodies against EGFR mutation proteins might be useful for the diagnosis and treatment of lung cancer.
authors.affiliation: Department of Pathology, The Central Hospital of Enshi Autonomous Prefecture, Enshi, Hubei, People's Republic of China.
authors.forename: Yan-Gang
authors.initials: YG
authors.lastname: Qu
authors.affiliation: Department of Molecular Pathology, Wuhan Nano Tumor Diagnosis Engineering Research Center, Wuhan, Hubei, People's Republic of China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Department of Thoracosurgery, Traditional Chinese Medical Hospital of Wenling, Wenling, Zhejiang, People's Republic of China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Pan
authors.affiliation: Department of Pathology, School of Basic Medical Science, Wuhan University, Wuhan, Hubei, People's Republic of China.
authors.forename: Xian-Da
authors.initials: XD
authors.lastname: Zhao
authors.affiliation: Department of Molecular Pathology, Wuhan Nano Tumor Diagnosis Engineering Research Center, Wuhan, Hubei, People's Republic of China.
authors.forename: Yan-Hua
authors.initials: YH
authors.lastname: Huang
authors.affiliation: Department of Thoracosurgery, Traditional Chinese Medical Hospital of Wenling, Wenling, Zhejiang, People's Republic of China.
authors.forename: Fu-Chun
authors.initials: FC
authors.lastname: Chen
authors.affiliation: Department of Pathology, School of Basic Medical Science, Wuhan University, Wuhan, Hubei, People's Republic of China.
authors.forename: Hong-Lei
authors.initials: HL
authors.lastname: Chen
mesh_terms: D000911:Antibodies, Monoclonal; D002289:Carcinoma, Non-Small-Cell Lung / Q000737:chemistry*; D066246:ErbB Receptors / Q000737:chemistry / Q000235:genetics*; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006801:Humans; D008175:Lung Neoplasms / Q000737:chemistry*; D009154:Mutation / Q000235:genetics; D045663:Quantum Dots / Q000737:chemistry; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D000911:Antibodies, Monoclonal; D066246:ErbB Receptors
keywords: EGFR; gene mutation; immunohistochemistry; lung cancer; quantum dots; real-time PCR