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使用基于自动试剂盒的PCR系统检测非小细胞肺癌患者血浆中的表皮生长因子受体(EGFR)突变

Detection of EGFR Mutations From Plasma of NSCLC Patients Using an Automatic Cartridge-Based PCR System.

作者信息

Heeke Simon, Hofman Véronique, Benzaquen Jonathan, Otto Josiane, Tanga Virginie, Zahaf Katia, Allegra Maryline, Long-Mira Elodie, Lassalle Sandra, Marquette Charles-Hugo, Ilie Marius, Hofman Paul

机构信息

Department of Thoracic H&N Medical Oncology, UT MD Anderson Cancer Center, Houston, TX, United States.

Laboratory of Clinical and Experimental Pathology, Biobank BB-0033-00025, Centre Hospitalier Universitaire Nice, Nice, France.

出版信息

Front Pharmacol. 2021 Apr 14;12:657743. doi: 10.3389/fphar.2021.657743. eCollection 2021.

Abstract

The introduction of liquid biopsies for the detection of mutations in non-small cell lung cancer patients (NSCLC) has revolutionized the clinical care. However, liquid biopsies are technically challenging and require specifically trained personnel. To facilitate the implementation of liquid biopsies for the detection of mutations from plasma, we have assessed a fully automated cartridge-based qPCR test that allows the automatic detection of mutations directly from plasma. We have analyzed 54 NSCLC patients and compared the results of the cartridge-base device to an FDA-approved assay. Detection of EGFR mutations was comparable but slightly lower in the cartridge-based device for L858R mutations (14/15 detected, 93%) and exon 19 deletions (18/20 detected, 90%). Unfortunately, 8/54 (15%) tests failed but increasing the proteinase K volume helped to recover 3/4 (75%) unsuccessful samples. In summary, the fully automated cartridge-based device allowed the detection of EGFR mutations directly from plasma in NSCLC patients with promising accuracy. However, protocol adjustments are necessary to reduce a high test failure rate.

摘要

液体活检用于检测非小细胞肺癌(NSCLC)患者的突变,这一技术的引入彻底改变了临床护理方式。然而,液体活检在技术上具有挑战性,需要经过专门培训的人员。为了便于实施通过血浆检测突变的液体活检技术,我们评估了一种基于盒式装置的全自动定量聚合酶链反应(qPCR)检测方法,该方法可直接从血浆中自动检测突变。我们分析了54例NSCLC患者,并将基于盒式装置的检测结果与一种经美国食品药品监督管理局(FDA)批准的检测方法的结果进行了比较。对于L858R突变(检测到14/15,93%)和外显子19缺失(检测到18/20,90%),基于盒式装置的EGFR突变检测结果相当,但略低。不幸的是,8/54(15%)的检测失败,但增加蛋白酶K的用量有助于挽救3/4(75%)未成功的样本。总之,基于盒式装置的全自动检测方法能够直接从NSCLC患者的血浆中检测EGFR突变,准确性令人满意。然而,需要对方案进行调整以降低高检测失败率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0037/8079969/7d2704520de7/fphar-12-657743-g001.jpg

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