Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: Production of orthoflavivirus recombinant NS1 for sensitive and specific microarray detection of orthoflavivirus IgG. ( a ) Recombinant NS1 and ( b ) recombinant EDIII of TBEV, West Nile virus (WNV) lineage 2, dengue virus (DENV) DENV 1 , DENV 2 , DENV 3 , DENV 4 , Zika virus (ZIKV), Japanese encephalitis virus (JEV) and Usutu virus (USUV) were purified and then analyzed via SDS-PAGE.

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Recombinant, Microarray, Virus, Purification, SDS Page

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: Overview of serum collection used for orthoflavivirus microarray validation.

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Microarray, Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Virus, Neutralization

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: Validation of orthoflavivirus recombinant NS1 for sensitive and specific microarray detection of orthoflavivirus IgG. Heatmaps displaying microarray titers for the orthoflavivirus rNS1 of ( a ) orthoflavivirus negative reference sera, ( b ) WNV patients, ( d ) ZIKV patients, ( f ) DENV patients and ( h ) TBEV patients. (The color key of the titers is indicated at the bottom of each heatmap). The microarray antigens are listed on the left of each heatmap. Violin plots showing microarray titers of ( c ) WNV patients, ( e ) ZIKV patients, ( g ) DENV patients and ( i ) TBEV natural infection-positive sera against the recombinant NS1 of TBEV, DENV1, DENV2, DENV3, DENV4, ZIKV, WNV, USUV and JEV. A non-parametric t -test was performed to assess the statistical differences of paired samples. Significance is noted as ***, p < 0.001; *, p < 0.5. ( b ) Receiving operating characterization (ROC) curve was created with GraphPad Prism software (version 9.1.0).

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Biomarker Discovery, Recombinant, Microarray, Infection, Software

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: ( a ) Heatmap displaying microarray titers for the orthoflavivirus rNS1 of orthoflavivirus TBEV-vaccinated patients. (The color key of the titers is indicated at the bottom of the heatmap). The microarray antigens are listed on the left of the heatmap. ( b ) Violin plot showing microarray titers of the pre-characterized TBEV, dengue virus (DENV), Zika virus (ZIKV), West Nile virus (WNV) sera and the negative orthoflavivirus sera against TBEV rEDIII. TBEV vaccinees were used as the positive control. ( c ) Heatmap displaying microarray titers for the orthoflavivirus rEDIII of orthoflavivirus TBEV-vaccinated patients. (The color key of the titers indicated at the bottom of the heatmap.) The microarray antigens are listed on the y-axis. ( d )Violin plot showing microarray titers of TBEV-vaccinated serum samples against the recombinant EDIII of TBEV, DENV1, DENV2, DENV3, DENV4, ZIKV, WNV, USUV and JEV. A non-parametric T-test was performed to assess the statistical differences of paired samples. Significance is noted as ****, p < 0.0001; **, p < 0.01; *, p < 0.5. The order of patient samples is the same for each heatmap.

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Microarray, Virus, Positive Control, Recombinant

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: TBEV-infected serum samples against recombinant EDIII. ( a ) Heatmap displaying microarray titers for the orthoflavivirus rEDIII of orthoflavivirus TBEV natural infection-positive sera. (The color key of the titers is indicated at the bottom of each heatmap). The microarray antigens are listed on the y-axis. ( b ) Violin plots showing the microarray titers of TBEV natural infection-positive sera against the recombinant EDIII of TBEV, DENV1, DENV2, DENV3, DENV4, ZIKV, WNV, USUV and JEV. A non-parametric T-test was performed to assess the statistical differences of paired samples. Significance is noted as ****, p < 0.0001; ***, p < 0.001; *, p < 0.5. ( c ) Receiving operating characterization (ROC) curve was created with GraphPad Prism software (version 9.1.0). The order of patient samples is the same as in .

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Infection, Recombinant, Microarray, Software

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: Evaluation of recombinant orthoflavivirus EDIII as an antigen. Heatmaps and violin plots showing the microarray titers of ZIKV-positive sera ( a , b ), DENV-positive sera ( c , d ) and WNV-positive sera ( e , f ) against the recombinant rEDIII of TBEV, DENV1, DENV2, DENV3, DENV4, ZIKV, WNV, USUV and JEV. The order of patient samples is the same as in .

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Recombinant, Microarray

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: All TBEV serology results of sera ( n = 5) that tested TBEV IgG positive for rNS1 and/or rEDIII in the orthoflavivirus microarray.

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Microarray, Enzyme-linked Immunosorbent Assay, Infection

Journal: Biotechnology Reports

Article Title: High-throughput microarray for antimicrobial susceptibility testing

doi: 10.1016/j.btre.2017.10.004

Figure Lengend Snippet: High-throughput AST chip. (A) A schematic of high-throughput antimicrobial susceptibility testing using AST-Chip. Antimicrobial susceptibility testing using the chip can be conducted in 12 h; Microarray micrograph of S. aureus cultures formed on hydrogel spots of AST-Chip after 12–18 h. The spots are interspaced at 1 mm with a spot diameter of 0.5 mm; The nano-biofilms analyzed using FUN-1 is demonstrated in green color; (B) BacLight live/dead assay shows metabolically active cells in yellow-orange color, present as colonies. The scale bar is 100 μm; (C) SEM micrograph of S. aureus nano-biofilm colonies. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Any subsequent dilutions needed for the antimicrobial susceptibility assays were made in PBS, mixed in a suspension of 0.5% alginate, and optimized media, and 50 nL of this mixture was spotted using a non-contact microarray spotter (Omnigrid Micro, Digilab Inc., Holliston, MA) on modified glass slides.

Techniques: High Throughput Screening Assay, Microarray, Live Dead Assay, Metabolic Labelling

Journal: Biotechnology Reports

Article Title: High-throughput microarray for antimicrobial susceptibility testing

doi: 10.1016/j.btre.2017.10.004

Figure Lengend Snippet: Antimicrobial susceptibility profile of S. aureus . AST of methicillin against wild-type (UAMS-1WT) and community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA). (A) Microarray scanner image of AST-Chip with S. aureus cultures exposed to different concentrations of methicillin [MET]. The viability is indicated by the fluorescence intensity of the spots. (B) Scanning electron micrographs of S. aureus biofilms that are untreated (top) or treated with 10 μg/ml MET (bottom); (C) Dose-response profile of S. aureus cultures obtained using AST-Chip.

Article Snippet: Any subsequent dilutions needed for the antimicrobial susceptibility assays were made in PBS, mixed in a suspension of 0.5% alginate, and optimized media, and 50 nL of this mixture was spotted using a non-contact microarray spotter (Omnigrid Micro, Digilab Inc., Holliston, MA) on modified glass slides.

Techniques: Microarray, Fluorescence