CAJ | 학술논문

目的：以MassARRAY质谱iPLEX分析技术为平台建立适合中国肺癌人群多基因同步检测的方法。方法：通过文献检索并结合国内外研究进展，确定与中国肺癌人群发病、耐药以及转移等密切相关的13个靶基因或相关传导通路基因（EGFR、KRAS、ALK、FGFR1、FGFR2、FGFR3、PIK3CA、BRAF、PTEN、MET、ERBB2、AKT1、STK11）。对目的基因突变进行筛选并确定99个热点突变。按MassARRAY突变位点标示方法和引物设计固定格式，引物设计软件在线设计297条引物（正、反向扩增引物和延伸引物各99条）。以8个肺癌细胞系以及6例肿瘤组织样本建立检测方法，与LungCarta试剂盒对比验证。扩大检测100例肺癌组织样本，与EGFR和KRAS直接测序法比较敏感度与特异度。结果：使用本方法检测肺癌细胞系的基因突变，其中1例肺癌组织新检测到FGFR2基因突变，其他结果与LungCarta试剂盒一致。与直接测序法相比较灵敏度为100%，特异度为96.3%。结论：成功建立MassARRAY质谱分析肺癌多基因突变检测方法，适合中国肺癌人群且具有临床应用前景。
목적：이MassARRAY질보iPLEX분석기술위평태건립괄합중국폐암인군다기인동보검측적방법。방법：통과문헌검색병결합국내외연구진전，학정여중국폐암인군발병、내약이급전이등밀절상관적13개파기인혹상관전도통로기인（EGFR、KRAS、ALK、FGFR1、FGFR2、FGFR3、PIK3CA、BRAF、PTEN、MET、ERBB2、AKT1、STK11）。대목적기인돌변진행사선병학정99개열점돌변。안MassARRAY돌변위점표시방법화인물설계고정격식，인물설계연건재선설계297조인물（정、반향확증인물화연신인물각99조）。이8개폐암세포계이급6례종류조직양본건립검측방법，여LungCarta시제합대비험증。확대검측100례폐암조직양본，여EGFR화KRAS직접측서법비교민감도여특이도。결과：사용본방법검측폐암세포계적기인돌변，기중1례폐암조직신검측도FGFR2기인돌변，기타결과여LungCarta시제합일치。여직접측서법상비교령민도위100%，특이도위96.3%。결론：성공건립MassARRAY질보분석폐암다기인돌변검측방법，괄합중국폐암인군차구유림상응용전경。
Objective:To establish a method based on the iPLEX analysis of MassARRAY mass spectrometry platform to detect multiplex genetic mutations among Chinese lung cancer patients. Methods:We reviewed the related literature and data of lung cancer treatments. We also determined 99 mutation hot spots in 13 target genes, namely, EGFR, KRAS, ALK, FGFR1, FGFR2, FGFR3, PIK3CA, BRAF, PTEN, MET, ERBB2, AKT1, and STK11, which are closely related to the pathogenesis, drug resistance, and metastasis of lung cancer and are associated with relevant transduction pathways. A total of 297 primers comprising 99 paired forward and reverse amplification primers and 99 matched extension primers were designed by using Assay Design in accordance with the mutation label and format requirements of the MassARRAY platform. The detection method was established by analyzing eight cell lines and six lung cancer specimens;the proposed method was then validated through comparisons with a LungCarta kit. The sensitivity and specificity of the proposed method were evaluated by directly sequencing EGFR and KRAS genes in 100 lung cancer cases. Results:The proposed method could detect multiplex genetic mutations in the lung cancer cell lines, and this finding is consistent with that observed using previously reported methods. The proposed method could also detect such mutations in clinical lung cancer specimens;this result is also consistent with that observed by using the LungCarta kit. However, an FGFR2 mutation was detected only by using the proposed method. The measured sensitivity and specificity were 100%and 96.3%, respectively. Conclusion:The proposed MassARRAY technology-based method could detect multiplex genetic mutations among Chinese lung cancer patients. Indeed, the proposed method can be potentially applied to detect mutations in cancer cells.