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新型基于PCR的全自动Idylla™ BRAF突变检测在恶性黑色素瘤福尔马林固定石蜡包埋组织上的多中心评估。

Multi-center evaluation of the novel fully-automated PCR-based Idylla™ BRAF Mutation Test on formalin-fixed paraffin-embedded tissue of malignant melanoma.

作者信息

Melchior Linea, Grauslund Morten, Bellosillo Beatriz, Montagut Clara, Torres Erica, Moragón Ester, Micalessi Isabel, Frans Johan, Noten Veerle, Bourgain Claire, Vriesema Renske, van der Geize Robert, Cokelaere Kristof, Vercooren Nancy, Crul Katrien, Rüdiger Thomas, Buchmüller Diana, Reijans Martin, Jans Caroline

机构信息

Department of Pathology, Copenhagen University Hospital, Copenhagen, Denmark.

Pathology Department, Hospital del Mar, Barcelona, Spain.

出版信息

Exp Mol Pathol. 2015 Dec;99(3):485-91. doi: 10.1016/j.yexmp.2015.09.004. Epub 2015 Sep 25.

Abstract

The advent of BRAF-targeted therapies led to increased survival in patients with metastatic melanomas harboring a BRAF V600 mutation (implicated in 46-48% of malignant melanomas). The Idylla(™) System (Idylla(™)), i.e., the real-time-PCR-based Idylla(™) BRAF Mutation Test performed on the fully-automated Idylla(™) platform, enables detection of the most frequent BRAF V600 mutations (V600E/E2/D, V600K/R/M) in tumor material within approximately 90 min and with 1% detection limit. Idylla(™) performance was determined in a multi-center study by analyzing BRAF mutational status of 148 archival formalin-fixed paraffin-embedded (FFPE) tumor samples from malignant melanoma patients, and comparing Idylla(™) results with assessments made by commercial or in-house routine diagnostic methods. Of the 148 samples analyzed, Idylla(™) initially recorded 7 insufficient DNA input calls and 15 results discordant with routine method results. Further analysis learned that the quality of 8 samples was insufficient for Idylla(™) testing, 1 sample had an invalid routine test result, and Idylla(™) results were confirmed in 10 samples. Hence, Idylla(™) identified all mutations present, including 7 not identified by routine methods. Idylla(™) enables fully automated BRAF V600 testing directly on FFPE tumor tissue with increased sensitivity, ease-of-use, and much shorter turnaround time compared to existing diagnostic tests, making it a tool for rapid, simple and highly reliable analysis of therapeutically relevant BRAF mutations, in particular for diagnostic units without molecular expertise and infrastructure.

摘要

BRAF靶向疗法的出现提高了携带BRAF V600突变的转移性黑色素瘤患者的生存率(46%-48%的恶性黑色素瘤与之相关)。Idylla™系统,即在全自动Idylla™平台上进行的基于实时PCR的Idylla™ BRAF突变检测,能够在约90分钟内检测肿瘤材料中最常见的BRAF V600突变(V600E/E2/D、V600K/R/M),检测限为1%。在一项多中心研究中,通过分析148例来自恶性黑色素瘤患者的存档福尔马林固定石蜡包埋(FFPE)肿瘤样本的BRAF突变状态,并将Idylla™结果与商业或内部常规诊断方法的评估结果进行比较,确定了Idylla™的性能。在分析的148个样本中,Idylla™最初记录了7个DNA输入不足的结果以及15个与常规方法结果不一致的结果。进一步分析发现,8个样本的质量不足以进行Idylla™检测,1个样本的常规检测结果无效,10个样本的Idylla™结果得到了确认。因此,Idylla™识别出了所有存在的突变,包括7个常规方法未识别出的突变。与现有诊断测试相比,Idylla™能够直接在FFPE肿瘤组织上进行全自动BRAF V600检测,具有更高的灵敏度、易用性和更短的周转时间,使其成为一种用于快速、简单且高度可靠地分析与治疗相关的BRAF突变的工具,尤其适用于没有分子专业知识和基础设施的诊断单位。

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