+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Serum and Plasma Copy Number Detection Using Real-time PCR



Serum and Plasma Copy Number Detection Using Real-time PCR



Journal of Visualized Experiments 2017(130):



Serum and plasma cell free DNA (cfDNA) has been shown as an informative, non-invasive source of biomarkers for cancer diagnosis, prognosis, monitoring, and prediction of treatment resistance. Starting from the hypothesis that androgen receptor (AR) gene copy number (CN) gain is a frequent event in metastatic castration resistance prostate cancer (mCRPC), we propose to analyze this event in cfDNA as a potential predictive biomarker. We evaluated AR CN in cfDNA using 2 different real-time PCR assays and 2 reference genes (RNaseP and AGO1). DNA amount of 60 ng was used for each assay combination. AR CN gain was confirmed using Digital PCR as a more accurate method. CN variation analysis has already been demonstrated to be informative for the prediction of treatment resistance in the setting of mCRPC, but it could be useful also for other purposes in different patient settings. CN analysis on cfDNA has several advantages: it is non-invasive, rapid and easy to perform, and it starts from a small volume of serum or plasma material.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 065506422

Download citation: RISBibTeXText

PMID: 29286483

DOI: 10.3791/56502


Related references

Real-Time PCR for the Detection of Precise Transgene Copy Number in Wheat. Methods in Molecular Biology 1679: 251-257, 2017

Real-time PCR for the detection of precise transgene copy number in durum wheat. Cellular and Molecular Biology Letters 16(4): 652-668, 2011

Comparison between real-time quantitative PCR detection of HER2 mRNA copy number in peripheral blood and ELISA of serum HER2 protein for determining HER2 status in breast cancer patients. Cellular Oncology 31(3): 203-211, 2009

Rapid and reliable detection of α-globin copy number variations by quantitative real-time PCR. Bmc Hematology 14(1): 4, 2014

Fast detection of MYCN copy number alterations in brain neuronal tumors by real-time PCR. Journal of Clinical Laboratory Analysis 22(2): 123-130, 2008

Quantitation of BCR-ABL transcript copy number after allogeneic bone marrow transplantation in patients with CML using real-time PCR detection. Blood 92(10 Suppl. 1 Part 1-2): 652A, 1998

Diagnosis of Pelizaeus-Merzbacher disease: detection of proteolipid protein gene copy number by real-time PCR. Neurogenetics 6(2): 73-78, 2005

TmRNA--a novel high-copy-number RNA diagnostic target--its application for Staphylococcus aureus detection using real-time NASBA. Fems Microbiology Letters 301(2): 218-223, 2009

Removal of Contaminant DNA by Combined UV-EMA Treatment Allows Low Copy Number Detection of Clinically Relevant Bacteria Using Pan-Bacterial Real-Time PCR. Plos one 10(7): E0132954, 2015

Real-time quantitative PCR as an alternative to Southern blot or fluorescence in situ hybridization for detection of gene copy number changes. Methods in Molecular Biology 353: 205-226, 2007

Detection of Stachybotrys chartarum using rRNA, tri5, and beta-tubulin primers and determining their relative copy number by real-time PCR. Mycological Research 112(Pt 7): 845-851, 2008

TaqMan-based real-time polymerase chain reaction for detection of FUT2 copy number variations: identification of novel Alu-mediated deletion. Transfusion 51(4): 762-769, 2011

Copy number of bean mitochondrial genes estimated by real-time PCR does not correlate with the number of gene loci and transcript levels. Plant Molecular Biology 61(1-2): 1-12, 2006

Detection of CAPN10 copy number variation in Thai patients with type 2 diabetes by denaturing high performance liquid chromatography and real-time quantitative polymerase chain reaction. Journal of Diabetes Investigation 6(6): 632-639, 2015

Real-time polymerase chain reaction technique determines absolute copy number of plasma methylated MGMT gene promoter copies in newly diagnosed malignant glioma patients. Journal of Clinical Oncology 26(15_Suppl): 22073-22073, 2016