Initial Diagnosis of ALK-Positive Non-Small-Cell Lung Cancer Based on Analysis of ALK Status Utilizing Droplet Digital PCR

详细信息    查看全文

• 作者：H. Louise Lund ; Curtis B. Hughesman ; Kareem Fakhfakh ; Kelly McNeil ; Shahira Clemens ; Kimberly Hocken ; Ryan Pettersson ; Aly Karsan ; Leonard J. Foster ; Charles Haynes
• 刊名：Analytical Chemistry
• 出版年：2016
• 出版时间：May 3, 2016
• 年：2016
• 卷：88
• 期：9
• 页码：4879-4885
• 全文大小：393K
• 年卷期：0
• ISSN：1520-6882

文摘

We describe a novel droplet digital PCR (ddPCR) assay capable of detecting genomic alterations associated with inversion translocations. It is applied here to detection of rearrangements in the anaplastic lymphoma kinase (ALK) gene associated with ALK-positive non-small-cell lung cancer (NSCLC). NSCLC patients may carry a nonreciprocal translocation on human chromosome 2, in which synchronized double stranded breaks (DSB) within the echinoderm microtubule-associated protein-like 4 (EML4) gene and ALK lead to an inversion of genetic material that forms the non-natural gene fusion EML4-ALK encoding a constitutively active tyrosine kinase that is associated with 3 to 7% of all NSCLCs. Detection of ALK rearrangements is currently achieved in clinics through direct visualization via a fluorescent in situ hybridization (FISH) assay, which can detect those rearrangements to a limit of detection (LOD) of ca. 15%. We show that the ddPCR assay presented here provides a LOD of 0.25% at lower cost and with faster turnaround times.