An In-Depth Evaluation of the Validity and Logistics Surrounding the Testing of AR-V7 mRNA Expression in Circulating Tumor Cells

Publication date: Available online 21 February 2018 Source:The Journal of Molecular Diagnostics Author(s): Anieta M. Sieuwerts, Bianca Mostert, Michelle van der Vlugt-Daane, Jaco Kraan, Corine Beaufort, Mai Van, Wendy J.C. Prager, Bram De Laere, Nick Beije, Paul Hamberg, Hans M. Westgeest, Metin Tascilar, Luc Y. Dirix, Wendy Onstenk, Ronald de Wit, Martijn P. Lolkema, Ron H.J. Mathijssen, John W.M. Martens, Stefan Sleijfer Recent reports have emphasized the clinical relevance of detecting the androgen receptor splice variant 7 (AR-V7) in circulating tumor cells (CTCs). Our aim was to set up a validated multicenter pipeline to measure AR-V7 by RT-qPCR in RNA isolated from CellSearch-enriched CTCs to provide an AR-V7 positive/negative score in a clinically acceptable time range. CellSearch-enirched CTCs from metastatic castration-resistant prostate cancer patients were characterized by RT-qPCR. After optimization it was prospectively tested whether it was possible to report the AR-V7 status within 11 days (PRELUDE study). In the range of the RNA equivalent of 0.2 to 12 VCaP cells the coefficient of variation for AR-V7 was 9% (n = 37). The LOD was 0.3 and the LOQ 3 cells in the final RT-qPCR. No differences were observed between AR-V7 data generated by four technicians or in two different laboratories. For the 45 patients in the PRELUDE study, 13 patients were ineligible, 22 patients were AR-V7 negative, and 10 AR-V7 positive. The median time to inform the...
Source: The Journal of Molecular Diagnostics - Category: Pathology Source Type: research