BEAMing and Droplet Digital PCR Analysis of Mutant IDH1 mRNA in Glioma Patient Serum and Cerebrospinal Fluid Extracellular Vesicles

被引：285

作者：

Chen, Walter W. ^{[1
,2
,3
,4
]}

Balaj, Leonora ^{[1
,2
,5
]}

Liau, Linda M. ^{[6
]}

Samuels, Michael L. ^{[7
]}

Kotsopoulos, Steve K. ^{[7
]}

Maguire, Casey A. ^{[1
,2
,3
]}

LoGuidice, Lori ^{[8
]}

Soto, Horacio ^{[6
]}

Garrett, Matthew ^{[6
]}

Zhu, Lin Dan ^{[1
,2
,3
]}

Sivaraman, Sarada ^{[1
,2
,3
]}

Chen, Clark ^{[9
,10
,11
]}

Wong, Eric T. ^{[9
,10
]}

Carter, Bob S. ^{[11
]}

Hochberg, Fred H. ^{[1
,2
,3
]}

Breakefield, Xandra O. ^{[1
,2
,3
]}

Skog, Johan ^{[8
]}

机构：

[1] Massachusetts Gen Hosp, Dept Neurol, Boston, MA 02114 USA

[2] Harvard Univ, Sch Med, Neurosci Program, Boston, MA 02115 USA

[3] Massachusetts Gen Hosp, Dept Radiol, Boston, MA 02114 USA

[4] Harvard Univ, Sch Med, Harvard Mit Div Hlth Sci & Technol, Boston, MA USA

[5] Amsterdam Canc Ctr, Neurooncol Res Grp, Amsterdam, Netherlands

[6] Univ Calif Los Angeles, David Geffen Sch Med, Med Ctr, Dept Neurosurg, Los Angeles, CA 90095 USA

[7] RainDance Technol, Lexington, MA USA

[8] Exosome Diagnost Inc, New York, NY USA

[9] Beth Israel Deaconess Med Ctr, Brain Tumor Ctr, Boston, MA 02215 USA

[10] Beth Israel Deaconess Med Ctr, Neurooncol Unit, Boston, MA 02215 USA

[11] Univ Calif San Diego, Dept Neurosurg, San Diego, CA 92103 USA

来源：

MOLECULAR THERAPY-NUCLEIC ACIDS | 2013年 / 2卷

关键词：

BEAMing PCR; biomarkers; cancer; extracellular vesicles; droplet digital PCR; INTEGRATED GENOMIC ANALYSIS; COLORECTAL-CANCER; GLIOBLASTOMA-MULTIFORME; CLINICAL-SAMPLES; TUMOR DNA; MUTATIONS; MICROVESICLES; BIOMARKERS; SEQUENCES; BREAST;

D O I：

10.1038/mtna.2013.28

中图分类号：

R-3 [医学研究方法]; R3 [基础医学];

学科分类号：

100103 [病原生物学]; 100218 [急诊医学];

摘要：

Development of biofluid-based molecular diagnostic tests for cancer is an important step towards tumor characterization and real-time monitoring in a minimally invasive fashion. Extracellular vesicles (EVs) are released from tumor cells into body fluids and can provide a powerful platform for tumor biomarkers because they carry tumor proteins and nucleic acids. Detecting rare point mutations in the background of wild-type sequences in biofluids such as blood and cerebrospinal fluid (CSF) remains a major challenge. Techniques such as BEAMing (beads, emulsion, amplification, magnetics) PCR and droplet digital PCR (ddPCR) are substantially more sensitive than many other assays for mutant sequence detection. Here, we describe a novel approach that combines biofluid EV RNA and BEAMing RT-PCR (EV-BEAMing), as well droplet digital PCR to interrogate mutations from glioma tumors. EVs from CSF of patients with glioma were shown to contain mutant IDH1 transcripts, and we were able to reliably detect and quantify mutant and wild-type IDH1 RNA transcripts in CSF of patients with gliomas. EV-BEAMing and EV-ddPCR represent a valuable new strategy for cancer diagnostics, which can be applied to a variety of biofluids and neoplasms.

引用

页数：10

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