4a330d1f839e62ae4546dc330d6a7443 pmc_33804266.pdf 67909c00ba3a53a1ca7ade4e6e7b839775be38fb pmc_33804266.pdf 17a53a6ef529c66d61becdba59a9b08cf5d99f7df791c7d2aab5f7c01634bbea pmc_33804266.pdf Title: Comparative Study of Gold and Carbon Nanoparticles in Nucleic Acid Lateral Flow Assay Subject: A lateral flow assay (LFA) is a paper-based, point-of-need test designed to detect a specific analyte in complex samples in low-resource settings. Although LFA has been successfully used in different applications, its use is still limited when high sensitivity is required, especially in the diagnosis of an early-stage condition. The limit of detection (LOD) is clearly related to the signal-generating system used to achieve the visual readout, in many cases involving nanoparticles coupled to a biomolecule, which, when combined, provides sensitivity and specificity, respectively. While colloidal gold is currently the most-used label, other detection systems are being developed. Carbon nanoparticles (CNPs) demonstrate outstanding features to improve the sensitivity of this technology by producing an increased contrast in the paper background. Based on the necessity of sensitivity improvement, the aim of this work is a comparative study, in terms of analytical performance, between commercial streptavidin gold nanoparticles (streptAv-AuNPs) and avidin carbon nanoparticles (Av-CNPs) in a nucleic acid lateral flow assay. The visual LOD of the method was calculated by serial dilution of the DNA template, ranging from 0.0 to 7 pg L-1/1.5 104 CFU mL-1). The LFA achieved visual detection of as low as 2.2 10-2 pg L-1 using Av-CNPs and 8.4 10-2 pg L-1 using streptAv-AuNPs. These LODs could be obtained without the assistance of any instrumentation. The results demonstrate that CNPs showed an increased sensitivity, achieving the nanomolar range even by visual inspection. Furthermore, CNPs are the cheapest labels, and the suspensions are very stable and easy to modify. Keywords: carbon black; gold nanoparticles; rapid diagnostic tests; lateral flow assay Author: Juan Carlos Porras, Mireia Bernuz, Jennifer Marfa, Arnau Pallares-Rusiñol, Mercè Martí and María Isabel Pividori Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Fri Mar 19 12:40:20 2021 CET ModDate: Fri Mar 19 12:48:09 2021 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 11 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3565846 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- PSUYZB+VnURWPalladioL Type 1 Custom yes yes no 10 0 HQUNPA+EURM10 Type 1 Builtin yes yes no 15 0 MEFMFE+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 HNPAQJ+URWPalladioL-Bold Type 1 Custom yes yes no 24 0 AKWXIG+URWPalladioL-Roma Type 1 Custom yes yes no 28 0 AWJYXN+CMSY10 Type 1 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PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 559 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Reagents and Equipment Destination: subsection.2.1 Item: Title: Carbon Nanoparticles Modification and Storage Destination: subsection.2.2 Item: Title: Characterization of CNPs by Transmission Electron Microscopy Destination: subsection.2.3 Item: Title: Bacterial Strains Culture, DNA Extraction, and Double-Tagging PCR Destination: subsection.2.4 Item: Title: Double-Tagging PCR and Quantification by Gel Electrophoresis Destination: subsection.2.5 Item: Title: Lateral Flow Assays Based on Carbon and Gold Nanoparticles as Signal-Generating Systems Destination: subsection.2.6 Item: Title: Data Interpretation and Analysis Destination: subsection.2.7 Item: Title: Results and Discussion Destination: section.3 Children: Item: Title: Characterization of CNPs by Transmission Electron Microscopy Destination: subsection.3.1 Item: Title: Double-Tagging PCR and Quantification by Gel Electrophoresis Destination: subsection.3.2 Item: Title: Lateral Flow Assays Based on Carbon and Gold Nanoparticles as Signal-Generating Systems Destination: subsection.3.3 Item: Title: Conclusions Destination: section.4 Item: Title: References Destination: section.4 Info: Title: Comparative Study of Gold and Carbon Nanoparticles in Nucleic Acid Lateral Flow Assay Author: Juan Carlos Porras, Mireia Bernuz, Jennifer Marfa, Arnau Pallares-Rusiñol, Mercè Martí and María Isabel Pividori Subject: A lateral flow assay (LFA) is a paper-based, point-of-need test designed to detect a specific analyte in complex samples in low-resource settings. Although LFA has been successfully used in different applications, its use is still limited when high sensitivity is required, especially in the diagnosis of an early-stage condition. The limit of detection (LOD) is clearly related to the signal-generating system used to achieve the visual readout, in many cases involving nanoparticles coupled to a biomolecule, which, when combined, provides sensitivity and specificity, respectively. While colloidal gold is currently the most-used label, other detection systems are being developed. Carbon nanoparticles (CNPs) demonstrate outstanding features to improve the sensitivity of this technology by producing an increased contrast in the paper background. Based on the necessity of sensitivity improvement, the aim of this work is a comparative study, in terms of analytical performance, between commercial streptavidin gold nanoparticles (streptAv-AuNPs) and avidin carbon nanoparticles (Av-CNPs) in a nucleic acid lateral flow assay. The visual LOD of the method was calculated by serial dilution of the DNA template, ranging from 0.0 to 7 pg L-1/1.5 104 CFU mL-1). The LFA achieved visual detection of as low as 2.2 10-2 pg L-1 using Av-CNPs and 8.4 10-2 pg L-1 using streptAv-AuNPs. These LODs could be obtained without the assistance of any instrumentation. The results demonstrate that CNPs showed an increased sensitivity, achieving the nanomolar range even by visual inspection. Furthermore, CNPs are the cheapest labels, and the suspensions are very stable and easy to modify. 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