single particle counting based on digital plasmonic nanobubble detection for rapid CORD-Papers-2021-10-25 (Version 1)

Title: Single-Particle Counting Based on Digital Plasmonic Nanobubble Detection for Rapid and Ultrasensitive Diagnostics
Abstract: Rapid and sensitive diagnostics of infectious diseases is an urgent and unmet need as evidenced by the COVID-19 pandemic. Here we report a novel strategy, based on DIgitAl plasMONic nanobubble Detection (DIAMOND), to address these gaps. Plasmonic nanobubbles are transient vapor bubbles generated by laser heating of plasmonic nanoparticles, and allow single-particle detection. Using gold nanoparticles labels and an optofluidic setup, we demonstrate that DIAMOND achieves a compartment-free digital counting and works on homogeneous assays without separation and amplification steps. When applied to the respiratory syncytial virus diagnostics, DIAMOND is 150 times more sensitive than commercial lateral flow assays and completes measurements within 2 minutes. Our method opens new possibilities to develop single-particle digital detection methods and facilitate rapid and ultrasensitive diagnostics.
Published: 2/23/2021
Journal: medRxiv : the preprint server for health sciences
DOI: 10.1101/2021.02.18.21252027
DOI_URL: http://doi.org/10.1101/2021.02.18.21252027
Author Name: Liu, Y
Author link: https://covid19-data.nist.gov/pid/rest/local/author/liu_y
Author Name: Ye, H
Author link: https://covid19-data.nist.gov/pid/rest/local/author/ye_h
Author Name: Huynh, H
Author link: https://covid19-data.nist.gov/pid/rest/local/author/huynh_h
Author Name: Kang, P
Author link: https://covid19-data.nist.gov/pid/rest/local/author/kang_p
Author Name: Xie, C
Author link: https://covid19-data.nist.gov/pid/rest/local/author/xie_c
Author Name: Kahn, J S
Author link: https://covid19-data.nist.gov/pid/rest/local/author/kahn_j_s
Author Name: Qin, Z
Author link: https://covid19-data.nist.gov/pid/rest/local/author/qin_z
sha: 94cc2ca9833c91b0c6c532dc2dc4087f7967495b
license: medrxiv
source_x: MedRxiv; Medline; WHO
source_x_url: https://www.medline.com/https://www.who.int/
pubmed_id: 33655274
pubmed_id_url: https://www.ncbi.nlm.nih.gov/pubmed/33655274
url: https://www.ncbi.nlm.nih.gov/pubmed/33655274/ http://medrxiv.org/cgi/content/short/2021.02.18.21252027v1?rss=1 https://doi.org/10.1101/2021.02.18.21252027
has_full_text: TRUE
Keywords Extracted from Text Content: COVID-19 subfemtomolar Single-particle serum plasMONic lateral core-satellites COVID-19 λ medRxiv sample-to-answer NPs AUC Synagis R 2 RSV ~333-fold bright/dark-field Fig. 3A λ=240 sub-femtomolar μ LFA kit medRxiv preprint lateral dashed lines Fig. S2, 3 compartment-free single-particle singleparticle SiO 2 DIAMOND RSV surface AuNPs μ+5 σ Palivizumab Fig. 4D capillary ~570fold particles' LFA AuNP ~150-fold NP nasal swab samples surface λ 5 children Fig. S1 3,3'-dithiobis Fig. 2C https://doi.org/10.1101/2021.02.18.21252027 doi DTSSP zone RSVspiked Fig. 4C plasMONic aqueous sulfosuccinimidyl propionate × PNBs
Extracted Text Content in Record: First 5000 Characters:Rapid and sensitive diagnostics of infectious diseases is an urgent and unmet need as evidenced by the COVID-19 pandemic. Here we report a novel strategy, based on DIgitAl plasMONic nanobubble Detection (DIAMOND), to address these gaps. Plasmonic nanobubbles are transient vapor bubbles generated by laser heating of plasmonic nanoparticles and allow single-particle 5 detection. Using gold nanoparticles labels and an optofluidic setup, we demonstrate that DIAMOND achieves a compartment-free digital counting and works on homogeneous assays without separation and amplification steps. When applied to the respiratory syncytial virus diagnostics, DIAMOND is 150 times more sensitive than commercial lateral flow assays and completes measurements within 2 minutes. Our method opens new possibilities to develop 10 single-particle digital detection methods and facilitate rapid and ultrasensitive diagnostics. One Sentence Summary: Single-particle digital plasmonic nanobubble detection allows rapid and ultrasensitive detection of viruses in a one-step homogeneous assay. , Single-molecule enzyme-linked 40 immunosorbent assay detects serum proteins at subfemtomolar concentrations. Nat. Biotechnol. 28, 595-599 (2010). The ability to rapidly detect diseases with high precision is of paramount importance as evidenced by the current COVID-19 pandemic (1, 2) . Digital assays have been a remarkable conceptual advance over the past two decades due to their single-molecule detection and absolute quantification (3, 4). They partition the analytes into microwells or emulsion droplets as 5 small compartments for independent signal amplification and digital counting, leading to the sensitivity enhancement by up to 10 3 -fold over the conventional assays (i.e., enzyme-linked immunosorbent assay and polymerase chain reaction) (3, 4) . Despite those advantages, digital assays have suffered from complex assay operations. Such paradigms prompt further innovations that develop various digital sensing platforms based on micro/nano-particles (5-7). With its 10 capability of examining individual particles' changes upon recognizing target molecules, singleparticle detection holds great potential to simplify digital assays (8-10). Examples of singleparticle digital assays include bright/dark-field imaging (11), interferometric (12) or fluorescent imaging (13, 14) , surface-enhanced Raman scattering (15), surface plasmon resonance microscopy imaging (16), and particle mobility tracking (17) (18) (19) . However, current techniques 15 rely on cumbersome particle purification and advanced imaging that inevitably limit their widespread use. Herein, we report a novel strategy for simplified single-particle digital assay based on DIgitAl plasMONic nanobubble Detection (DIAMOND, see Table S1 for comparison with other assays). Plasmonic nanobubbles (PNBs) refer to the vapor bubbles generated by short laser pulse 20 excitation of plasmonic nanoparticles (NPs) and amplify their intrinsic scattering for the detection by a secondary probe laser (20- 24) . They have lifetime that lasts nanoseconds and are sensitive to the physical parameters of NPs such as sizes, shape, concentration, and clustering state (20-22). Taking advantage of these unique properties, we designed an optofluidic setup to flow the NP suspension in a micro-capillary ( Fig. 1A and Fig. S1 ). The focused laser beam 25 probes a microscale "virtual compartment" of about 16 pL and detects the PNB generation from single particles. Since PNBs are transient events, there is no cross-talk between laser pulses, which allows "on" and "off" signal counting in a compartment-free manner (Fig. 1B) . We demonstrated robust single NP counting and sensitive detection of large particles in a strong background of small particles (1 in 240). We then implemented DIAMOND in a homogeneous 30 assay that uses plasmonic gold NPs (AuNPs) as labels without additional separation or amplification steps. Using silica (SiO 2 ) beads as targets, we demonstrated that DIAMOND has sub-femtomolar detection limit and provides absolute quantification. When applied to detect respiratory syncytial virus (RSV), DIAMOND provided a 150-fold sensitivity enhancement over the state-of-the-art lateral flow assay (LFA) and sample-to-answer time within 2 minutes. 35 Therefore, DIAMOND opens new possibilities to develop separation-, amplification-, and compartment-free single-particle digital assays and facilitate rapid and ultrasensitive diagnostic platforms. We first evaluated the ability of DIAMOND for single NP detection. Serial aqueous dilutions 40 of 75 nm AuNP (as characterized in Fig. S2, 3 and Table S2 ) were prepared for the DIAMOND tests. For simplicity, we converted the particle concentration into the expected average number (λ) of AuNPs per detection zone. Fig. 2A and Fig. S4 show the representative testing results and the corresponding Poisson distributions for the given λ , respectively. Due to the variation of actual AuNP nu
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