Rapid Isolation and Detection for RNA Biomarkers for TBI Diagnostics
Technical Report,30 Sep 2015,29 Sep 2016
University of California, San Diego La Jolla United States
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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.
- Medicine and Medical Research