Rapid Isolation and Detection for RNA Biomarkers for TBI Diagnostics
Abstract
Our project work is focused on using a new dielectrophoresis (DEP) microarray technology for rapid isolation and detection of brain-specific RNA and other biomarkers for TBI diagnostics and patient monitoring. Initial year one project goals include developing DEP techniques for isolation of circulating cell-free (ccf) RNA from glioblastoma exosomes and TBI samples (IRB dependent); methods for on-chip fluorescence detection of exosomes, RNA, and other biomarkers; methods for isolation of mRNA and miRNA from exosomes; and development of RT-PCR techniques and specific primers for identifying brain-specific mRNAs, miRNAs and other biomarkers. We have been successful in demonstrating DEP isolation of glioblastoma exosomes from 50 of un-diluted plasma in fifteen to twenty minutes. We also showed tri-color fluorescent detection of the isolated exosomes (red fluorescence), ccf-RNA (green fluorescence) and ccf-DNA (blue fluorescence). Finally, glioblastoma-specific mRNAs for EGFRvIII as well as more universal beta-actin biomarkers were identified by RT-PCR carried out on ccf-RNA extracted from the exosomes.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2015
- Accession Number
- AD1002695
Entities
People
- Michael J Heller
Organizations
- University of California, San Diego