MassARRAY, pyrosequencing, and PNA clamping for EGFR mutation detection in lung cancer tissue and cytological samples: a multicenter study.

Background: Testing for epidermal growth factor receptor (EGFR) mutation is an important process in the therapeutic plan of patients with lung cancer. Recently, MassARRAY, based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, has been shown to be a useful method for somatic mutation analysis with pyrosequencing and peptide nucleic acid clamping (PNAc).

Methods: A total of 107 tissues and 67 cytological samples, which were confirmed to have lung adenocarcinoma at nine hospitals in Korea, were collected. Among the MassARRAY, pyrosequencing, and PNAc, the concordance rates and sensitivity of EGFR mutation detection were analyzed and validated in comparative tissue and cytological specimens.

Results: The concordance rate between pyrosequencing and PNAc was higher than that between MassARRAY and either of the pyrosequencing and PNAc in both tissue and cytological samples. In a comparison of diagnostic performance, MassARRAY (sensitivity: 85.7 %) was higher than pyrosequencing (74.3 %) and PNAc (70 %) in tissue, although pyrosequencing (80.5 %) was more highly sensitive, compared to MassARRAY (70.7 %) and PNAc (70.7 %) in terms of cytology. Unexpectedly, use of MassARRAY resulted in a significantly different EGFR mutation detection rate between tissue and cytological samples.

Conclusions: When used for the detection of EGFR mutations, MassARRAY was more sensitive than pyrosequencing or PNA clamping in tissue, but not in cytological samples. In EGFR mutation detection between tissues and cytology, PNAc showed relatively higher concordance than MassARRAY or pyrosequencing.