Simultaneous Detection of Exosomal Membrane Protein and RNA by Highly Sensitive Aptamer Assisted Multiplex–PCR
journal contribution
posted on 2019-11-11, 16:12 authored by Zaizai Dong, Chuanhao Tang, Zhen Zhang, Wei Zhou, Rong Zhao, Lina Wang, Jiachao Xu, Yayun Wu, Jiang Wu, Xing Zhang, Li Xu, Libo Zhao, Xiaohong FangExosomes,
which are 30–150 nm extracellular vesicles containing
a variety of biomolecules (i.e., proteins, RNA, DNA, and lipids),
have emerged as important analytes in liquid biopsy. Although exosome
detection offers a valuable chance to understand disease status in
a multidimensional manner, comprehensive analysis of different exosomal
biomolecules with limited specimens remains a challenge. Herein, we
introduced an aptamer-based PCR (polymerase chain reaction) platform
and realized highly sensitive detection of CD63 protein positive exosomes
as low as 50 particles/μL. More importantly, simultaneous analysis
of exosomal RNA and surface protein is available using this method.
In a proof of concept experiment, simultaneous detection of two immunotherapy-related
biomarkers, programmed death-ligand 1 (PD-L1) protein and indoleamine
2,3-dioxygenase 1 (IDO1) mRNA, was demonstrated with clinical samples.
Our method provides a strategy for the comprehensive analysis of exosomes,
with the advantages of high sensitivity and practicability.
