Establishment and application of a multiplex genetic mutation-detection method of lung cancer based on MassARRAY platform

Hong-Xia Tian, Xu-Chao Zhang, Zhen Wang, Jian-Guang Chen, Shi-Liang Chen, Wei-Bang Guo, Yi-Long Wu

Abstract


Objective: This study aims to establish a method for highly parallel multiplexed detection of genetic mutations in Chinese lungcancer samples through Agena iPLEX chemistry and matrix-assisted laser desorption ionization time-of-flight analysis onMassARRAY mass spectrometry platform.

Methods: We reviewed the related literature and data on lung cancer treatments. We also identified 99 mutation hot spots in 13target genes closely related to the pathogenesis, drug resistance, and metastasis of lung cancer. A total of 297 primers, composed of99 paired forward and reverse amplification primers and 99 matched extension primers, were designed using Assay Designsoftware. The detection method was established by analyzing eight cell lines and six lung cancer specimens. The proposed methodwas then validated through comparisons by using a LungCartaTM kit. The sensitivity and specificity of the proposed method wereevaluated by directly sequencing EGFR and KRAS genes in 100 lung cancer cases.

Results: The proposed method was able to detect multiplex genetic mutations in lung cancer cell lines. This finding was consistentwith the observations on previously reported mutations. The proposed method can also detect such mutations in clinical lungcancer specimens. This result was consistent with the observations with LungCartaTM kit. However, an FGFR2 mutation wasdetected only through the proposed method. The measured sensitivity and specificity were 100% and 96.3%, respectively.

Conclusions: The proposed MassARRAY technology-based multiplex method can detect genetic mutations in Chinese lung cancerpatients. Therefore, the proposed method can be applied to detect mutations in other cancer tissues.


Keywords


Lung neoplasms; driver genes; mutation; multigene testing; MassARRAY

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