RET基因重排的检测在甲状腺乳头状癌的评价与优化方法的建议-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 Zhongsheng Sun,2 Qunsheng Ji,1 Guanshan Zhu,1 and Xinying Su1* 1Asia Emerging MarketsiMed,AstraZeneca RD.199 LiangJing Road,ZhangJiang Hi-Tech Park,Shanghai 201203,China 2Institute 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), ?uorescence in situ hybridization (FISH), and immunohistochemistry (IHC). RET/PTC rearrangement was detected using either RET break-apart FISH followed by multi- color FISH to con?rm 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 speci?city and sensitivity of multiplex qPCR and IHC were calculated using break-apart FISH as a reference. Among 73 PTC patients with suf?cient tissue available for FISH and multiplex qPCR, 10 cases were de?ned 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 identi?ed another two CCDC6/RET fusion positive cases, but with low RET mRNA expression. IHC staining identi?ed 11 RET positive cases among 39 patients with available samples. In comparison to FISH, multiplex qPCR displayed 100% sensitivity and 97% speci?city to detect RET/PTC fusions, while IHC was neither sensitive nor speci?c. Our data reveal that both multiplex qPCR