RET基因重排的检测在甲状腺乳头状癌的评价和优化方法的建议-E.pdfVIP

GENES, CHROMOSOMES CANCER 00:00–00 (2014) RESEARCH ARTICLE An Evaluation and Recommendation of the Optimal Methodologies to Detect RET Gene Rearrangements in Papillary Thyroid Carcinoma Tianwei Zhang, 1 Yachao Lu, 1 Qingqing Ye, 1 Meizhuo Zhang, 1 Li Zheng, 1 Xiaolu Yin, 1 Paul Gavine, 1 2 1 1 1 Zhongsheng Sun, Qunsheng Ji, Guanshan Zhu, and Xinying Su * 1 Asia Emerging MarketsiMed, AstraZeneca RD.199 LiangJing Road, ZhangJiang Hi-Tech Park, Shanghai 201203,China 2 Institute of Genomic Medicine,Wenzhou Medical University, Wenzhou, Zhejiang 325000,China To recommend a reliable and clinically realistic RET/PTC rearrangement detection assay for papillary thyroid carcinoma (PTC), we compared multiplex quantitative polymerase chain reaction (qPCR), fluorescence in situ hybridization (FISH), and immunohistochemistry (IHC). RET/PTC rearrangement was detected using either RET break-apart FISH followed by multi- color FISH to confirm CCDC6/RET or NCOA4/RET fusions, or by multiplex qPCR to detect 14 RET/PTC subtypes with simul- taneous RET mRNA expression. RET protein expression was detected by IHC. The specificity and sensitivity of multiplex qPCR and IHC were calculated using break-apart FISH as a reference. Among 73 PTC patients with sufficient tissue available for FISH and multiplex qPCR, 10 cases were defined as RET/PTC positive by both assays, including eight CCDC6/RET and two NCOA4/RET fusions with relatively high RET mRNA. In addition, multiplex qPCR identified another two CCDC6/RET fusion positive cases, but with low RET mRNA expression. IHC staining identified 11 RET positive cases among 39 patients with available sam