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). Materials and 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). Conclusion 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.

Applications and Immunological Effects of Quantum Dots on Respiratory System

“…Su et al reported iodine doped carbon dots conjugated with cetuximab as a dual fluorescent/CT probe for bioimaging lung cancer cells epidermal growth factor receptor (EGFR) (16). Additionally, QDs immunofluorescence histochemistry (QDs-IHC) was employed to detect EGFR mutant, RRM2 and Bcl2, and Monocarboxylate transporter 4 (MCT-4) in non-small cell lung cancer patients (17).…”

Emerging trends in the novel drug delivery approaches for the treatment of lung cancer

“…During in-vivo studies, QDs coated with PEG were not recognized by the reticuloendothelial system increasing their circulation time within the biological system [123]. QDs are also used for biological imaging, cellular and molecular tracing, estimating toxicity profile, in-vitro imaging (multiplexed), molecular labelling as markers, signal transduction and immuno-histochemical detection [124]. The aqueous solubility of QDs can be increased by coating them with polymer and ligand exchange [71] [125].…”

Quantum Dot Based Nano-Biosensors for Detection of Circulating Cell Free miRNAs in Lung Carcinogenesis: From Biology to Clinical Translation

“…Furthermore, designing probes to detect disease specific mutations is an attractive approach toward early clinical identification of the different cancers. QDs linked to EGFR mutation-specific antibodies were assessed and reported to be highly effective and sensitive as compared with traditional methods in disease diagnosis and therapeutic decision making for NSCLC patients (Qu et al, 2014). In an attempt to further improve the diagnostic ability of these molecules, a highly specific QDs based recognition-before-labeling strategy using DNA aptamers was developed.…”