Abstract:

Objective To establish a rapid screening method for epidermal growth factor receptor（EGFR） gene L858R mutation in peripheral blood by loop-mediated isothermal amplification（LAMP） among non-small cell lung cancer（NSCLC） patients. Methods The DNA sequence of exon 21（L858R） of EGFR gene was searched in Genebank,and then the Primer Explorer V4 software was used to design specific primers. LAMP was established,and the specificity and sensitivity were evaluated. Peripheral plasma of 11 patients with NSCLC diagnosed by clinical pathology was collected,and the accuracy of LAMP was verified using the amplification results of digital polymerase chain reaction（PCR） as standard. Results EGFR gene L858R mutant DNA can be specifically amplified by LAMP primer and reaction system designed by ourselves. The detection sensitivity was 0.1 %,and the specificity reached 100%. LAMP detected 9 positive cases in 11 NSCLC specimens with EGFR gene L858R mutation positive results. Conclusions The LAMP has been established for detecting EGFR gene mutation in human peripheral plasma,which has high sensitivity,specificity and accuracy. It can be used to visually interpret the detection results by fluorescence. It is a simple and rapid screening method for EGFR gene mutation.

Key words: Loop-mediated isothermal amplification, Epidermal growth factor receptor, Circulating tumor DNA, Point-of-care testing