Journal: Trends in analytical chemistry : TRAC

Article Title: Ultraspecific analyte detection by direct kinetic fingerprinting of single molecules

doi: 10.1016/j.trac.2019.115764

Figure Lengend Snippet: Nucleic acid detection technologies and their analytical performance.

Article Snippet: Bio-Rad CFX96) Extraction kits, primers, fluorogenic probes, thermostable polymerase, dNTPs ddPCR [ 27 , 33 ] Yes Amplification-based zM to fM 99.99% Limited Droplet generator + droplet reader (e.g., Bio-Rad QX100), thermal cycler Extraction kits, droplet generation reagents, primers, fluorogenic probes, thermostable polymerase Illumina sequencing (TruSeq) [ 27 ] Yes Amplification-based zM to fM >99% High Illumina sequencer (e.g., MiSeq, HiSeq, NovaSeq), thermal cycler Extraction kits, library preparation kits, fluorescent dNTPs, thermostable polymerase, flow cells Microarray [ 27 , 31 ] Yes Amplification-free nM to mM 50–80% High Microarray plate scanner (e.g., AffyMetrix GeneChip HT) Extraction and PCR/labeling kits, microarray chips ADNA [ 22 ] Yes Amplification-free aM to fM Not available Not available Dark field microscope (e.g., Zeiss Axiovert 200M) Probe-conjugated nanoparticles, patterned lipid arrays NanoString [ 27 ] Yes Amplification-free fM to nM 90–95% High nCounter system (e.g., nCounter MAX ) Flow cells, capture probes, fluorescently barcoded affinity probes Simoa [ 24 ] Yes Amplification-free fM to pM >95% Limited Simoa analyzer (e.g., HD-1, SR-X) Probe-conjugated microparticles, droplet generation reagents, primary and secondary detection probes, enzymatic signal developing reagents Time gated FRET ligation [ 26 ] Yes Amplification-free pM to nM Not available Limited Fluorescence plate reader (e.g., ThermoFisher KRYPTOR) FRET probes, adaptor probes, DNA or RNA ligase SiMREPS [ 17 ] Yes Amplification-free fM to pM ≥99.99999% Limited TIRF microscope (e.g., Olympus IX83, Oxford NanoImager) Flow cells, capture probes, fluorescent probes Open in a separate window Nucleic acid detection technologies and their analytical performance.

Techniques: Amplification, Multiplexing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Sequencing, Microarray, Microscopy, Ligation, Fluorescence

Journal: Toxins

Article Title: A High-Throughput, Precipitating Colorimetric Sandwich ELISA Microarray for Shiga Toxins

doi: 10.3390/toxins6061855

Figure Lengend Snippet: Standard curve for colorimetric sandwich ELISA microarray detection of Stx1 and/or Stx2. Well bottoms of a 96-well microtiter plate were contact printed (eight replicates) from left to right with an HRP-antibody marker (Column 1), two different monoclonal antibodies (mAbs) to Stx1 (1-2 in Column 2 and 1-1 in Column 3) and a mAb raised against Stx2 (2-2 in Column 4). Serial dilutions of Stx1, Stx2 and Stx1 mixed with Stx2 were reacted and visualized with the addition of a cocktail of the mAbs (HRP-labeled) followed by colorimetric development with an HRP-precipitating substrate. A representative sample flatbed scanned image of Stx1 and Stx2 detection at 125, 62, 31 and 0 ng/mL (top to bottom) before ( a ) and after ( b ) the addition of diluted (1:3 in distilled water) skin lotion. The standard curves for the detection of Stx1 ( c ); Stx2 ( d ); and Stx1 mixed with Stx2 ( e ).

Article Snippet: Colorimetric scans of the microarrayed microtiter plates were acquired with a Spotware Colorimetric Microarray scanner (Arrayit Corp., Sunnyvale, CA, USA).

Techniques: Sandwich ELISA, Microarray, Marker, Bioprocessing, Labeling

Journal: Toxins

Article Title: A High-Throughput, Precipitating Colorimetric Sandwich ELISA Microarray for Shiga Toxins

doi: 10.3390/toxins6061855

Figure Lengend Snippet: The colorimetric ELISA microarray detection of Stx produced by enriched axenic broth cultures of STECs. Various strains of STECs and non-STEC controls were cultured overnight in TSB ± mitomycin C to observe the antibiotic induction of toxin formation, as detected by a sandwich ELISA microarray platform with colorimetric detection. B-PER, added to a portion of aliquots from overnight cultures, was statically incubated or shaken during incubation, and samples were tested for the release of cell-associated toxin. The stationary or final cell concentration was determined via total aerobic plate culture with TSA. (The mitomycin C concentration was 50 ng/mL; the capture antibody was 1:2 diluted; Color development reactions were for 60 min prior to scanning.) Legend: (?) questionable Shiga toxin detection (changed to “+” when allowed to react overnight); (−) no discernable detection; (+) low detection response; (++) moderate detection response; (+++) high detection response.

Article Snippet: Colorimetric scans of the microarrayed microtiter plates were acquired with a Spotware Colorimetric Microarray scanner (Arrayit Corp., Sunnyvale, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Microarray, Produced, Cell Culture, Sandwich ELISA, Incubation, Concentration Assay

Journal: Toxins

Article Title: A High-Throughput, Precipitating Colorimetric Sandwich ELISA Microarray for Shiga Toxins

doi: 10.3390/toxins6061855

Figure Lengend Snippet: Colorimetric ELISA microarray detection of Stx produced by enriched broth cultures of STECs containing ground beef. Various strains of STECs and non-STEC controls were cultured overnight in TSB ± mitomycin C to observe the antibiotic induction of toxin formation as detected by a sandwich ELISA microarray platform with colorimetric detection. B-PER, added to a portion of aliquots from overnight cultures, was directly added to overnight cell culture aliquots held in the microtiter plate wells and statically incubated to promote the release of cell-associated toxin. (Inocula were 77 ± 40 CFU/mL; capture antibody was 1:10 diluted; the mitomycin C concentration was 66 ng/mL; color development reactions were for 60 min prior to scanning.) Legend: (?) questionable Shiga toxin detection (changed to “+” when allowed to react overnight); (−) no discernable detection; (+) low detection response; (++) moderate detection response; (+++) high detection response.

Article Snippet: Colorimetric scans of the microarrayed microtiter plates were acquired with a Spotware Colorimetric Microarray scanner (Arrayit Corp., Sunnyvale, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Microarray, Produced, Cell Culture, Sandwich ELISA, Incubation, Concentration Assay