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 protein biomarkers for TBI diagnostics and patient monitoring. Initial year one project goals include developing DEP techniques for isolation of cell-free (cf) RNA from glioblastoma exosomes and TBI samples (IRB dependent); methods for on-chip fluorescence detection of exosomes, RNA, and protein 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 protein biomarkers. We have been successful in demonstrating DEP isolation of glioblastoma exosomes from 50 L 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). Glioblastoma-specific EGFRvIII mRNA from exosomes isolated by DEP, as well as more universal beta-actin mRNA biomarkers, were identified by RT-PCR. We have now also developed a novel on-chip method to carryout immunofluorescence analysis of brain specific exosomal protein biomarkers. We believe this new technique is very close to achieving true Sample to Answer diagnostics.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2016
- Accession Number
- AD1033785
Entities
People
- Michael J Heller
Organizations
- University of California, San Diego