INV-17081
Extracellular vesicles are secreted by cells and contain essential biological components such as proteins, lipids, and genetic materials. Cells use these vesicles to communicate with other cells. These vesicles could be used to introduce therapeutic agents in cells and detect the diseased condition but, detecting cell-secreted extracellular vesicles (40-100nm diameter) is challenging with existing techniques. Current detection methods include the use of highly specific, complex, and expensive instruments. An additional challenge is that large quantities of exosomes are required for detection by ELISA; a time-consuming process. Hence, there is a need for a novel detection method of exosomes.
The invention proposes the use of highly sensitive and specific Rolling Circle Amplification (RCA) to multiplex detection of cell-specific extracellular vesicles in microfluidic droplets.
Researchers have developed a microfluidic method in which oligonucleotide probes and RCA reaction mixture are co-encapsulated for qualitative and quantitative detection of the secretions. These exosomes can be distinguished by selecting a panel of markers and multiplexing detection of several markers to specifically identify exosomes from each cell population. RCA is highly advantageous as it offers flexibility to recognize multiple targets on the exosomes by hybridizing oligonucleotides which may or may not be coupled to antibodies, depending on the identification of mRNA, miRNA, or protein markers. This can provide an accelerated and accurate detection method.