Aptamer-Initiated Catalytic Hairpin Assembly Fluorescence Assay for Universal, Sensitive Exosome Detection

Abstract

Cancer-cell-derived exosomes are regarded as noninvasive biomarkers for early cancer diagnosis because of their critical roles in intercellular communication and molecular exchange. A robust aptamer-initiated catalytic hairpin assembly (AICHA) fluorescence assay is proposed for universal, sensitive detection of cancer-derived exosomes. The AICHA was verified with the specific detection of MCF-7 cell-derived exosomes with a wide calibration range of 8.4 particles/μL to 8.4 × 105 particles/μL and a low detection limit (LOD) of 0.5 particles/μL. The universality of the AICHA method was verified for PANC-1 cell-derived exosomes, the LOD of which was determined to be 0.1 particles/μL. The performances in serum samples were detected with a recovery rate range of 95.45–106.2%, which demonstrates its significant potential for protein biomarker analysis and cancer diagnosis.

Recent Advances in Biomolecular Detection Based on Aptamers and Nanoparticles

“…Recently, catalytic hairpin assembly (CHA) was used as an enzyme-free technique for signal amplification with high sensitivity and selectivity. Zhou et al [ 83 ] proposed the detection of exosomes using aptamer-initiated CHA (AICHA) fluorescence signal amplification, as shown in Figure 7 D. In this strategy, target-specific aptamers hybridized with an initiator and conjugated to the surface of MNPs. In the presence of exosomes, the aptamers could recognize and bind to the exosomes due to the higher affinity between the aptamers and membrane proteins of exosomes, resulting in the release of initiators into the supernatant.…”

“…Numbers 1–7 represent the 7 steps used in the procedure. ( D ) AICHA signal amplification strategy for exosome detection, reprinted with permission from [ 83 ]. Copyright 2022 American Chemistry Society.…”

Clustered Regularly Interspaced Short Palindromic Repeats-Associated Proteins13a combined with magnetic beads, chemiluminescence and reverse transcription-recombinase aided amplification for detection of avian influenza a (H7N9) virus

“…And incompatible reaction systems often need to combine microfluidic or manually open the lid to take samples from the first step reaction, which is not conducive to clinical large-scale sample screening. ( Peng et al, 2020 ; Wang et al, 2020 ; Zhou et al, 2022 ).…”

Localized DNA tetrahedrons assisted catalytic hairpin assembly for the rapid and sensitive profiling of small extracellular vesicle-associated microRNAs

“…However, the clinical application of RT-qPCR is greatly impeded due to the tediousness of operation, specialized experimental equipment required, and high false positive rate [10,11]. In this regard, multiple methods have been proposed for sEV-miRNA analysis, such as the use of nanomaterial-based technologies [12][13][14], enzymeassisted strategies [15][16][17], and spherical nucleic acid [18][19][20]. Despite their decent performances, these strategies are not rapid and simple enough for sEV-miRNAs measurement because of the tedious operation and harsh reaction conditions.…”

“…Accordingly, metastable hairpindriven amplification, including catalytic hairpin assembly (CHA) [21][22][23], hybridization chain reaction (HCR) [24,25], have been proposed due to its high specificity, lowcost, and easy to use. Compared with the assembled concatemer products of HCR, the dangled ssDNA products of CHA are easier to capture and generate an enhanced electrochemical signal [18,26,27]. However, the deficient sensitivity and inefficiency hindered the practical application of metastable hairpin-driven amplification.…”