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Analytical platform evaluation for quantification of ERG in prostate cancer using protein and mRNA detection methods

Overview of attention for article published in Journal of Translational Medicine, February 2015
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Title
Analytical platform evaluation for quantification of ERG in prostate cancer using protein and mRNA detection methods
Published in
Journal of Translational Medicine, February 2015
DOI 10.1186/s12967-015-0418-z
Pubmed ID
Authors

Jintang He, Athena A Schepmoes, Tujin Shi, Chaochao Wu, Thomas L Fillmore, Yuqian Gao, Richard D Smith, Wei-Jun Qian, Karin D Rodland, Tao Liu, David G Camp, Anshu Rastogi, Shyh-Han Tan, Wusheng Yan, Ahmed A Mohamed, Wei Huang, Sreedatta Banerjee, Jacob Kagan, Sudhir Srivastava, David G McLeod, Shiv Srivastava, Gyorgy Petrovics, Albert Dobi, Alagarsamy Srinivasan

Abstract

The established methods for detecting prostate cancer (CaP) are based on tests using PSA (blood), PCA3 (urine), and AMACR (tissue) as biomarkers in patient samples. The demonstration of ERG oncoprotein overexpression due to gene fusion in CaP has thus provided ERG as an additional biomarker. Based on this, we hypothesized that ERG protein quantification methods can be of use in the diagnosis of prostate cancer. An antibody-free assay for ERG3 protein detection was developed based on PRISM (high-pressure high-resolution separations with intelligent selection and multiplexing)-SRM (selected reaction monitoring) mass spectrometry. We utilized TMPRSS2-ERG positive VCaP and TMPRSS2-ERG negative LNCaP cells to simulate three different sample types (cells, tissue, and post-DRE urine sediment). Enzyme-linked immunosorbent assay (ELISA), western blot, NanoString, and qRT-PCR were also used in the analysis of these samples. Recombinant ERG3 protein spiked into LNCaP cell lysates could be detected at levels as low as 20 pg by PRISM-SRM analysis. The sensitivity of the PRISM-SRM assay was approximately 10,000 VCaP cells in a mixed cell population model of VCaP and LNCaP cells. Interestingly, ERG protein could be detected in as few as 600 VCaP cells spiked into female urine. The sensitivity of the in-house ELISA was similar to the PRISM-SRM assay, with detection of 30 pg of purified recombinant ERG3 protein and 10,000 VCaP cells. On the other hand, qRT-PCR exhibited a higher sensitivity, as TMPRSS2-ERG transcripts were detected in as few as 100 VCaP cells, in comparison to NanoString methodologies which detected ERG from 10,000 cells. Based on this data, we propose that the detection of both ERG transcriptional products with RNA-based assays, as well as protein products of ERG using PRISM-SRM assays, may be of clinical value in developing diagnostic and prognostic assays for prostate cancer given their sensitivity, specificity, and reproducibility.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 23 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 23 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 8 35%
Professor 3 13%
Student > Ph. D. Student 3 13%
Student > Master 2 9%
Other 1 4%
Other 2 9%
Unknown 4 17%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 6 26%
Agricultural and Biological Sciences 4 17%
Medicine and Dentistry 3 13%
Computer Science 2 9%
Psychology 1 4%
Other 1 4%
Unknown 6 26%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 2. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 12 February 2015.
All research outputs
#14,215,235
of 22,789,076 outputs
Outputs from Journal of Translational Medicine
#1,780
of 3,988 outputs
Outputs of similar age
#190,598
of 357,813 outputs
Outputs of similar age from Journal of Translational Medicine
#47
of 115 outputs
Altmetric has tracked 22,789,076 research outputs across all sources so far. This one is in the 35th percentile – i.e., 35% of other outputs scored the same or lower than it.
So far Altmetric has tracked 3,988 research outputs from this source. They typically receive a lot more attention than average, with a mean Attention Score of 10.5. This one has gotten more attention than average, scoring higher than 50% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 357,813 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 44th percentile – i.e., 44% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 115 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 50% of its contemporaries.