Journal: ACS Nano

Article Title: Separation-Free Extracellular Vesicle Microribonucleic Acid Profiling Using Structurally Oriented Membrane Fusion and Spatially Confined Amplification

doi: 10.1021/acsnano.5c18632

Figure Lengend Snippet: Schematic Representation of DBS-Alip Selectively Fusing with EVs to Deliver a Dual-Color Biosensor, Enabling Rapid, Separation-Free Detection of EV miRNAs with Amplified Fluorescence for Early NSCLC Diagnosis (Figure Created with Biorender.com)

Article Snippet: Primers and template cDNA were mixed with all-in-one miRNA qRT-PCR detection kit mixture (GeneCopoeia) for amplification.

Techniques: Amplification, Fluorescence, Biomarker Discovery

Journal: ACS Nano

Article Title: Separation-Free Extracellular Vesicle Microribonucleic Acid Profiling Using Structurally Oriented Membrane Fusion and Spatially Confined Amplification

doi: 10.1021/acsnano.5c18632

Figure Lengend Snippet: In situ detection of miRNA-21/205 in NSCLC-associated EVs. (A, B) TCGA database was used to analyze the expression of miR-21/205 in lung squamous cell carcinoma and lung adenocarcinoma samples and normal samples. (C) qPCR evaluation of miR-21/205 expression in A549 and HBE cell-derived EVs, respectively. miR-21 Data were analyzed by Unpaired t test (** p < 0.01). miR-205 Data were analyzed by Unpaired t test with Welch’s correction (** p < 0.01). Data were presented as the means ± SEM, n = 3. (D) Selective detection of miR-21 and miR-205 within A549 and HBE EV vesicles using DBS-Alip. Data were analyzed by Mann–Whitney U test (** p < 0.01). Data were presented as the means ± SEM, n = 3. (E, F) Plot of the intensities of DBS detection signals in DBS-Alip vs the spiked concentration of EV. Data were presented as the means ± SEM, n = 3.

Article Snippet: Primers and template cDNA were mixed with all-in-one miRNA qRT-PCR detection kit mixture (GeneCopoeia) for amplification.

Techniques: In Situ, Expressing, Derivative Assay, MANN-WHITNEY, Concentration Assay

Journal: ACS Nano

Article Title: Separation-Free Extracellular Vesicle Microribonucleic Acid Profiling Using Structurally Oriented Membrane Fusion and Spatially Confined Amplification

doi: 10.1021/acsnano.5c18632

Figure Lengend Snippet: Validation of a multiple EV miRNA in situ detection method for lung cancer diagnosis. (A) Schematic representation of the targeted fusion-mediated multiple EV miRNA in situ detection method for distinguishing nonsmall cell lung cancer (NSCLC) from healthy individuals. Figure created with Biorender.com . (B) Heatmap of risk scores for differentially indicated markers and miRNA panels in 22 advanced NSCLC patients and 30 healthy controls. (C, D) Box plots illustrating the expression of markers that differentiate NSCLC patients from healthy controls. Boxes span 25th–75th percentiles, median indicated by horizontal line. Whiskers extend to minimum and maximum values, with all data points overlaid as individual dots. Data were analyzed by Mann–Whitney U test (**** p < 0.0001). (E) Receiver operating characteristic (ROC) curves (left) and area under the curve (AUC) values (right) for individual markers or marker combinations in discriminating NSCLC patients from healthy controls. (F) Confusion matrix demonstrating an overall diagnostic accuracy of 92.3% for marker combinations in differentiating NSCLC patients from healthy controls.

Article Snippet: Primers and template cDNA were mixed with all-in-one miRNA qRT-PCR detection kit mixture (GeneCopoeia) for amplification.

Techniques: Biomarker Discovery, In Situ, Expressing, MANN-WHITNEY, Marker, Diagnostic Assay

Journal: ACS Nano

Article Title: Separation-Free Extracellular Vesicle Microribonucleic Acid Profiling Using Structurally Oriented Membrane Fusion and Spatially Confined Amplification

doi: 10.1021/acsnano.5c18632

Figure Lengend Snippet: Validation of Cohorts from different regions for early Lung Cancer Diagnosis. (A) Heatmap displaying the abundance of various marker miRNAs and combination models in samples from eight healthy individuals and 12 early-stage NSCLC patients. The schematic portion of A was created with BioRender.com . (B, C) Box plots illustrating the use of individual markers to differentiate between early-stage NSCLC patients and those with healthy controls. Boxes span 25th–75th percentiles, median indicated by horizontal line. Whiskers extend to minimum and maximum values, with all data points overlaid as individual dots. Data were analyzed by Mann–Whitney U test (** p < 0.01, *** p < 0.001). (D) ROC analysis for the identification of healthy individuals and early-stage NSCLC patients using individual markers or marker combinations. (E) Heatmap displaying the abundance of various marker miRNAs and combination models in samples from twenty-one patients with benign pneumonia and eight additional early-stage NSCLC patients. The schematic portion of panel (E) was created with BioRender.com . (F, G) Box plots illustrating the use of individual markers to differentiate between early-stage NSCLC patients and those with benign pneumonia controls. Boxes span 25th–75th percentiles, median indicated by horizontal line. Whiskers extend to minimum and maximum values, with all data points overlaid as individual dots. Data were analyzed by Mann–Whitney U test (** p < 0.01). (H) ROC analysis for the identification of benign pneumonia patients and early-stage NSCLC patients using individual markers or marker combinations.

Article Snippet: Primers and template cDNA were mixed with all-in-one miRNA qRT-PCR detection kit mixture (GeneCopoeia) for amplification.

Techniques: Biomarker Discovery, Marker, MANN-WHITNEY