PMID- 25407564
OWN - NLM
STAT- MEDLINE
DCOM- 20150923
LR  - 20191210
IS  - 1098-2264 (Electronic)
IS  - 1045-2257 (Linking)
VI  - 54
IP  - 3
DP  - 2015 Mar
TI  - An evaluation and recommendation of the optimal methodologies to detect RET gene 
      rearrangements in papillary thyroid carcinoma.
PG  - 168-76
LID - 10.1002/gcc.22229 [doi]
AB  - 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 multicolor FISH to confirm CCDC6/RET or NCOA4/RET 
      fusions, or by multiplex qPCR to detect 14 RET/PTC subtypes with simultaneous 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 samples. In comparison to FISH, 
      multiplex qPCR displayed 100% sensitivity and 97% specificity to detect RET/PTC 
      fusions, while IHC was neither sensitive nor specific. Our data reveal that both 
      multiplex qPCR and FISH assays are equally applicable for detection of RET/PTC 
      rearrangements. Break-apart FISH methodology is highly recommended for the wider 
      screening of RET rearrangements (regardless of partner genes), while multiplex 
      qPCR is preferred to identify all known fusion types using one assay, provided 
      mRNA expression is also measured. IHC analysis could potentially provide an 
      additional method of fusion detection dependent on further optimization of assay 
      conditions and scoring cutoffs.
CI  - (c) 2014 Wiley Periodicals, Inc.
FAU - Zhang, Tianwei
AU  - Zhang T
AD  - Asia & Emerging Markets iMed, AstraZeneca R&D. 199 LiangJing Road, ZhangJiang 
      Hi-Tech Park, Shanghai, 201203, China.
FAU - Lu, Yachao
AU  - Lu Y
FAU - Ye, Qingqing
AU  - Ye Q
FAU - Zhang, Meizhuo
AU  - Zhang M
FAU - Zheng, Li
AU  - Zheng L
FAU - Yin, Xiaolu
AU  - Yin X
FAU - Gavine, Paul
AU  - Gavine P
FAU - Sun, Zhongsheng
AU  - Sun Z
FAU - Ji, Qunsheng
AU  - Ji Q
FAU - Zhu, Guanshan
AU  - Zhu G
FAU - Su, Xinying
AU  - Su X
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
DEP - 20141119
PL  - United States
TA  - Genes Chromosomes Cancer
JT  - Genes, chromosomes & cancer
JID - 9007329
RN  - EC 2.7.10.1 (Proto-Oncogene Proteins c-ret)
RN  - EC 2.7.10.1 (RET protein, human)
SB  - IM
MH  - Carcinoma/*genetics
MH  - Carcinoma, Papillary
MH  - Humans
MH  - Immunohistochemistry
MH  - *In Situ Hybridization, Fluorescence
MH  - Proto-Oncogene Proteins c-ret/*genetics
MH  - *Real-Time Polymerase Chain Reaction
MH  - Thyroid Cancer, Papillary
MH  - Thyroid Neoplasms/*genetics
MH  - *Translocation, Genetic
EDAT- 2014/11/20 06:00
MHDA- 2015/09/24 06:00
CRDT- 2014/11/20 06:00
PHST- 2014/07/16 00:00 [received]
PHST- 2014/10/22 00:00 [revised]
PHST- 2014/11/03 00:00 [accepted]
PHST- 2014/11/20 06:00 [entrez]
PHST- 2014/11/20 06:00 [pubmed]
PHST- 2015/09/24 06:00 [medline]
AID - 10.1002/gcc.22229 [doi]
PST - ppublish
SO  - Genes Chromosomes Cancer. 2015 Mar;54(3):168-76. doi: 10.1002/gcc.22229. Epub 
      2014 Nov 19.