应用酶切富集PCR法检测非小细胞肺癌患者胸腔积液表皮生长因子受体基因突变

Detection of EGFR Exon 19 and 21 Mutations in Pleural Effusion from Non-Small-Cell Lung Cancer Patients by Mutant Enriched PCR Assay

目的探讨酶切富集PCR法检测非小细胞肺癌(NSCLC)患者胸腔积液表皮生长因子受体基因(EGFR)外显子19缺失和外显子21 L858R突变的临床意义。方法采用EGFR基因突变酶切富集及非酶切富集PCR法对30例NSCLC患者胸腔积液游离核酸EGFR基因外显子19缺失和外显子21 L858R突变进行分析。结果30例NSCLC患者中,酶切富集PCR法分别检出EGFR基因外显子19缺失10例(33.3%)和外显子21 L858R突变5例(1.7%),非酶切富集PCR法则仅能分别检出6例(20.0%)和1例(3.3%);两种方法检出率差异有统计学意义(P=0.032)。在接受吉非替尼治疗的4例患者中,2例检出EGFR基因突变患者治疗后肿瘤均部分缓解。结论酶切富集PCR法可以高效、经济、准确地检测NSCLC患者胸腔积液游离核酸EGFR基因突变,可能成为临床NSCLC患者选择EGFR酪氨酸激酶抑制剂治疗的预测方法。

Objective To investigate the feasibility of detection of epidermal growth factor receptor （EGFR） exon 19 deletions and exon 21 L858R mutations in pleural effusion from non-small-cell lung cancer （NSCLC） patients by mutant enriched PCR assay. Methods The mutations of exon 19 and 21 of EGFR gene in pleural samples from thirty NSCLC patients were analyzed using both the mutant-enriched PCR assay and the non-enriched PCR assay. Results Ten （33.3%, 10/30） exon 19 deletions and five （ 16. 7%, 5/30） exon 21 L858R mutation were detected by the mutant-enriched PCR assay, while only 6 cases （20. 0% ） and 1 case （3.3%） were detected by the non-enriched PCR assay respectively. The difference of mutation detection rate of EGFR gene between the two methods was statistically significant （ P = 0. 032 ）. Mutations were detected in all of partial responders （2/4） among the four patients who received gefitinib therapy. Conclusions Mutant-enriched PCR assay can detect EGFR exon 19 deletions and exon 21 L858R mutation in pleural effusion from NSCLC patients effectively, economically and accurately. It may be a valuable biomarker for gefitinib therapy in advanced NSCLC.