Journal: Chemical Science

Article Title: Highly specific and rapid glycan based amperometric detection of influenza viruses † †Electronic supplementary information (ESI) available: Synthesis and characterization of all compounds and intermediates are provided. See DOI: 10.1039/c6sc03720h Click here for additional data file.

doi: 10.1039/c6sc03720h

Figure Lengend Snippet: Workflow and structure of the glycans used for the electrochemical detection. Top: electrochemical detection of influenza and S. pneumoniae . Bottom: structure of the glycans used for the electrochemical assay.

Article Snippet: It was gratifying to observe the viral strains, H1N1 (A/Brisbane/59/2007) and H3N2 (A/Victoria/361/2011) cleave all four compounds extremely well; in contrast, the two strains of S. pneumoniae (serotype 1, ATCC 6301 and serotype 1, ATCC 6305) cleave only the natural substrates, ( ) and not (4,7di-OMe)Sα2,3Gal or (4,7di-OMe)Sα2,6Gal.

Techniques:

Journal: Chemical Science

Article Title: Highly specific and rapid glycan based amperometric detection of influenza viruses † †Electronic supplementary information (ESI) available: Synthesis and characterization of all compounds and intermediates are provided. See DOI: 10.1039/c6sc03720h Click here for additional data file.

doi: 10.1039/c6sc03720h

Figure Lengend Snippet: Detection of NA using (A) natural substrates Sα2,3Gal and Sα2,6Gal or (B) new substrates (4,7di-OMe)Sα2,3Gal and (4,7di-OMe)Sα2,6Gal. 50 U of different enzymes [(1) H5N1 NA (A/Anhui/1/2005), (2) H3N2 NA (A/Babol/36/2005), (3) NA from Streptococcus pneumoniae specific for α2-3 linkages, (4) NA from Salmonella typhimurium specific for α2-3 linkages, (5) NA from Clostridium perfringens specific for α2-3, α2-6 and α2-8 linkages and (6) NA from Arthrobacter ureafaciens specific for α2-3, α2-6, α2-8 and α2-9 linkages] was incubated with Sα2,3Gal or Sα2,6Gal at 37 °C for 1 h. (C) Detection of influenza and S. pneumoniae . Different substrate [(Sα2,3Gal:Sα2,6Gal; (4,7di-OMe)Sα2,3Gal or (4,7di-OMe)Sα2,6Gal)] was incubated with different pathogens [H1N1 (A/Brisbane/59/2007); H3N2 (A/Victoria/361/2011); SP1 (serotype 1, ATCC 6301) or SP2 (serotype 1, ATCC6305)] at 37 °C for 1 h. (D) Monitoring antiviral efficacy. 10 ng of Zanamivir was premixed with different pathogens [(H3N2 A/Victoria/361/2011, VZ ), SP2 (serotype 1, ATCC 6305, SZ ), or H3N2 A/Victoria/361/2011 and SP2 ( VSZ )] for 30 min before the addition of Sα2,3Gal. Also shown is data obtained without the addition of Zanamivir to the pathogens [(H3N2 A/Victoria/361/2011, V ), SP2 (serotype 1, ATCC 6305, SP2 ), or A/Victoria/361/2011 and SP2 ( VS )]. The current was measured using Accu-Chek Aviva strips (Roche Diagnostics) with the 10 s average current recorded 5 s following substrate introduction at a working potential of –0.15 V. The y axis, Δ I , represents the difference in current before and after the addition of the substrate. All experiments were performed in triplicate independently on different days to demonstrate scientific rigor.

Article Snippet: It was gratifying to observe the viral strains, H1N1 (A/Brisbane/59/2007) and H3N2 (A/Victoria/361/2011) cleave all four compounds extremely well; in contrast, the two strains of S. pneumoniae (serotype 1, ATCC 6301 and serotype 1, ATCC 6305) cleave only the natural substrates, ( ) and not (4,7di-OMe)Sα2,3Gal or (4,7di-OMe)Sα2,6Gal.

Techniques: Incubation

Journal:

Article Title: Sensitive and Specific Method for Rapid Identification of Streptococcus pneumoniae Using Real-Time Fluorescence PCR

doi: 10.1128/JCM.39.10.3446-3451.2001

Figure Lengend Snippet: Serial dilutions of S. pneumoniae (ATCC 33400) DNA isolated by a standardized method and quantitated spectrophotometrically to 4.4e6 through 4.4e0 genomic equivalents were used for determination of real-time PCR assay detection limits or in vitro sensitivity testing. Cycle number plotted against the log of calculated concentration values resulted in a standard curve with an error of 0.592 and correlation coefficient at unity. Human genomic DNA at 4,500 genomic equivalents and NTC samples did not fluoresce above background signal. The detection limits of the PCR assay demonstrated similar results when the dilution series panel was run in testing of all cross-reaction panel and unknown organisms.

Article Snippet: The detection limits of the PCR assay demonstrated similar results when the dilution series panel was run in testing of all cross-reaction panel and unknown organisms. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Genus Species or serovar(s) (subtypes) Homo H. sapiens Streptococcus S. agalactiae, S. bovis, S. equi, S. equisimilis, S. pyogenes, S. sanguis (type II) Campylobacter C. coli, C. lari, C. jejuni Citrobacter C. freundii Escherichia E. coli H1, O28, O55, O111, 112, 0126, 0128, O157:H7 Klebsiella K. pneumoniae Leclercia L. adecarboxylata Neisseria N. lactamica Proteus P. vulgaris Pseudomonas P. aeruginosa Salmonella Bovis-morbificus, Choleraesuis, Cubana, enteritidis, Heidelberg, Infantis, Javiana, Lanka, Kovka, Montevideo, Newport, Paratyphi-A, Poona Typhi-1 (1078), Typhimurium (528) Shigella S. flexneri, S. boydii (type I) S. dysenteriae (type III) Staphylococcus S. aureus Mycoplasma M. pneumoniae, M. hominis Open in a separate window Cross-reactivity panel: negative-control organisms In addition, 1.0 ng of genomic DNA from laboratory stock, as well as DNA purified by the modified capture disk method, from each of 10 S. pneumoniae strains (ATCC 6305, ATCC 49619, ATCC 33400, ATCC 51915, 1301, 1346, 1518, 1661, 1830, and 2113) were correctly identified by the PCR assay.

Techniques: Isolation, Real-time Polymerase Chain Reaction, In Vitro, Concentration Assay

Journal:

Article Title: Sensitive and Specific Method for Rapid Identification of Streptococcus pneumoniae Using Real-Time Fluorescence PCR

doi: 10.1128/JCM.39.10.3446-3451.2001

Figure Lengend Snippet: Cross-reactivity panel: negative-control organisms

Article Snippet: The detection limits of the PCR assay demonstrated similar results when the dilution series panel was run in testing of all cross-reaction panel and unknown organisms. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Genus Species or serovar(s) (subtypes) Homo H. sapiens Streptococcus S. agalactiae, S. bovis, S. equi, S. equisimilis, S. pyogenes, S. sanguis (type II) Campylobacter C. coli, C. lari, C. jejuni Citrobacter C. freundii Escherichia E. coli H1, O28, O55, O111, 112, 0126, 0128, O157:H7 Klebsiella K. pneumoniae Leclercia L. adecarboxylata Neisseria N. lactamica Proteus P. vulgaris Pseudomonas P. aeruginosa Salmonella Bovis-morbificus, Choleraesuis, Cubana, enteritidis, Heidelberg, Infantis, Javiana, Lanka, Kovka, Montevideo, Newport, Paratyphi-A, Poona Typhi-1 (1078), Typhimurium (528) Shigella S. flexneri, S. boydii (type I) S. dysenteriae (type III) Staphylococcus S. aureus Mycoplasma M. pneumoniae, M. hominis Open in a separate window Cross-reactivity panel: negative-control organisms In addition, 1.0 ng of genomic DNA from laboratory stock, as well as DNA purified by the modified capture disk method, from each of 10 S. pneumoniae strains (ATCC 6305, ATCC 49619, ATCC 33400, ATCC 51915, 1301, 1346, 1518, 1661, 1830, and 2113) were correctly identified by the PCR assay.

Techniques: Negative Control

Journal:

Article Title: Sensitive and Specific Method for Rapid Identification of Streptococcus pneumoniae Using Real-Time Fluorescence PCR

doi: 10.1128/JCM.39.10.3446-3451.2001

Figure Lengend Snippet: Results of double blind PCR-based testing of clinical isolates

Article Snippet: The detection limits of the PCR assay demonstrated similar results when the dilution series panel was run in testing of all cross-reaction panel and unknown organisms. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Genus Species or serovar(s) (subtypes) Homo H. sapiens Streptococcus S. agalactiae, S. bovis, S. equi, S. equisimilis, S. pyogenes, S. sanguis (type II) Campylobacter C. coli, C. lari, C. jejuni Citrobacter C. freundii Escherichia E. coli H1, O28, O55, O111, 112, 0126, 0128, O157:H7 Klebsiella K. pneumoniae Leclercia L. adecarboxylata Neisseria N. lactamica Proteus P. vulgaris Pseudomonas P. aeruginosa Salmonella Bovis-morbificus, Choleraesuis, Cubana, enteritidis, Heidelberg, Infantis, Javiana, Lanka, Kovka, Montevideo, Newport, Paratyphi-A, Poona Typhi-1 (1078), Typhimurium (528) Shigella S. flexneri, S. boydii (type I) S. dysenteriae (type III) Staphylococcus S. aureus Mycoplasma M. pneumoniae, M. hominis Open in a separate window Cross-reactivity panel: negative-control organisms In addition, 1.0 ng of genomic DNA from laboratory stock, as well as DNA purified by the modified capture disk method, from each of 10 S. pneumoniae strains (ATCC 6305, ATCC 49619, ATCC 33400, ATCC 51915, 1301, 1346, 1518, 1661, 1830, and 2113) were correctly identified by the PCR assay.

Techniques: Negative Control