Abstract:

Objective To explore a new method for detection of epidermal grouth factor receptor (EGFR) mutations based on suspension array technology, which can be used to detect and analyse EGFR mutations associated with EGFR tyrosine kinase inhibitors (TKI) in tissues of non-small cell lung cancer (NSCLC). Methods Sixty-one patients with NSCLC confirmed by imageology and pathology were selected, and the fresh tumor tissues were collected after operation for DNA extraction. The multiplex amplifications of exons 18-21 containing 8 EGFR mutations occurring in high frequency were performed by polymerase chain reaction (PCR), followed by ligation of the PCR products with a series of special probes using ligase detection reaction (LDR), then the LDR products were detected by suspension array technology platform. The results were compared with that from direct sequencing for confirming EGFR mutations. Results Nineteen EGFR mutations in 61 tissue samples of NSCLD (31.1%) were detected using multiplex PCR-LDR combined with suspension array technology, including 1 E746-A750 del (1) deletion mutation and 1 E746-A750 del (2) deletion mutation in exon 19, 1 T790M point mutation in exon 20, and 14 L858R point mutation and 2 L861Q point mutation in exon 21. These EGFR mutations were verified by direct sequencing. Conclusion A new method for detection of EGFR mutations with high throughput and specificity has been established by PCR-LDR combined with suspension array technology, which can be used for accurate identification of 8 EGFR mutations occurring in high frequency simultaneously, and might be useful in detection of EGFR mutations for patients with NSCLC prior to targeted therapy with EGFR-TKI.

Key words: epidermal growth factor receptor, suspension array technology, polymerase chain reaction, ligase detection reaction, non-small cell lung cancer