PMID- 21808235
OWN - NLM
STAT- MEDLINE
DCOM- 20111122
LR  - 20230216
IS  - 1530-0307 (Electronic)
IS  - 0023-6837 (Linking)
VI  - 91
IP  - 10
DP  - 2011 Oct
TI  - High-throughput detection of fusion genes in cancer using the Sequenom MassARRAY 
      platform.
PG  - 1491-501
LID - 10.1038/labinvest.2011.110 [doi]
AB  - Fusion genes have pivotal roles in the development and progression of human 
      cancer and offer potential for rational drug design. Massively parallel 
      sequencing has identified a panoply of in-frame expressed fusion genes, but early 
      reports suggest that the majority of these are present at very low prevalence or 
      are private events. Conventional methods for the identification of recurrent 
      expressed fusion genes in large cohorts of cancers (eg fluorescence in situ 
      hybridization (FISH) and reverse transcriptase PCR (RT-PCR)) are time consuming 
      and prone to artifacts. Here, we describe a novel high-throughput strategy for 
      the detection of recurrent fusion genes in cancer based on the Sequenom MassARRAY 
      platform. Fusion genes were initially identified by massively parallel sequencing 
      of breast cancer cell lines. For each fusion gene, two Sequenom probes were 
      designed. Primary human breast cancers and cancer cell lines were interrogated 
      for 10 fusion genes. Sensitivity, specificity, and predictive values of the 
      MassARRAY method were then determined using FISH and qRT-PCR as the 'gold 
      standard.' By combining two probes per fusion gene, the negative and positive 
      predictive values were 100 and 71.4%, respectively. All fusion genes identified 
      by massively parallel sequencing were accurately detected. No recurrent fusion 
      genes were found. The MassARRAY-based approach described here may, therefore, be 
      employed as a high-throughput screening tool for known fusion genes in human 
      cancer. In keeping with other highly sensitive assays, further refinement of this 
      technique is necessary to reduce the number of false-positive results.
CI  - (c) 2011 USCAP, Inc All rights reserved
FAU - Lambros, Maryou B K
AU  - Lambros MB
AD  - Molecular Pathology Team, The Breakthrough Breast Cancer Research Centre, The 
      Institute of Cancer Research, London, UK.
FAU - Wilkerson, Paul M
AU  - Wilkerson PM
FAU - Natrajan, Rachael
AU  - Natrajan R
FAU - Patani, Neill
AU  - Patani N
FAU - Pawar, Vidya
AU  - Pawar V
FAU - Vatcheva, Radost
AU  - Vatcheva R
FAU - Mansour, Marthe
AU  - Mansour M
FAU - Laschet, Mirja
AU  - Laschet M
FAU - Oelze, Beatrice
AU  - Oelze B
FAU - Orr, Nicholas
AU  - Orr N
FAU - Muller, Susanne
AU  - Muller S
FAU - Reis-Filho, Jorge S
AU  - Reis-Filho JS
LA  - eng
GR  - BREAST CANCER NOW RESEARCH CENTRE/BCN_/Breast Cancer Now/United Kingdom
GR  - G1100450/MRC_/Medical Research Council/United Kingdom
GR  - WT_/Wellcome Trust/United Kingdom
GR  - CRUK_/Cancer Research UK/United Kingdom
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110801
PL  - United States
TA  - Lab Invest
JT  - Laboratory investigation; a journal of technical methods and pathology
JID - 0376617
RN  - 0 (Oncogene Proteins, Fusion)
SB  - IM
MH  - Breast Neoplasms/*genetics
MH  - Cell Line, Tumor
MH  - Female
MH  - Genetic Testing/standards
MH  - HeLa Cells
MH  - High-Throughput Nucleotide Sequencing/*methods/standards
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Microarray Analysis/*methods/standards
MH  - *Oncogene Proteins, Fusion
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sensitivity and Specificity
EDAT- 2011/08/03 06:00
MHDA- 2011/12/13 00:00
CRDT- 2011/08/03 06:00
PHST- 2011/08/03 06:00 [entrez]
PHST- 2011/08/03 06:00 [pubmed]
PHST- 2011/12/13 00:00 [medline]
AID - S0023-6837(22)02716-7 [pii]
AID - 10.1038/labinvest.2011.110 [doi]
PST - ppublish
SO  - Lab Invest. 2011 Oct;91(10):1491-501. doi: 10.1038/labinvest.2011.110. Epub 2011 
      Aug 1.