Journal: PLoS Pathogens

Article Title: Characterizing Emerging Canine H3 Influenza Viruses

doi: 10.1371/journal.ppat.1008409

Figure Lengend Snippet: Viruses used in the study and their abbreviations.

Article Snippet: After incubating in block solution (2.5% BSA in PBS) for 1 hour at room temperature, cells were incubated with the following mAbs (1μg/ml) or pAbs (dilution 1:1000) diluted in blocking solution for 1 hour at 37°C: anti-HA A/duck/Shantou/1283/2001 H3N8 goat pAb (BEI Resources NR-34586), A/equine/Miami/1/1963 H3N8 mouse mAb equine 7.1, or A/Hiroshima/52/2005 mouse pAb (BEI Resources F-287) for the staining of rCIV H3N2-, rCIV H3N8- or r/Wy H3N2-infected cells, respectively; anti-NA A/Singapore/1/1957 H2N2 goat pAb (BEI Resources NR-3137), A/equine/Miami/1/1963 H3N8 goat pAb (BEI Resources NR-3145) or hmAb 109219 (kindly provided by James J. Kobie) for the staining of rCIV H3N2-, rCIV H3N8- or r/Wy H3N2-infected cells, respectively; anti-NS1 1A7and 4D4 [ ] mouse mAbs cocktail (1:1, 2 μg/ml).

Techniques: Virus

Journal: PLoS Pathogens

Article Title: Characterizing Emerging Canine H3 Influenza Viruses

doi: 10.1371/journal.ppat.1008409

Figure Lengend Snippet: The presence of antibodies against rCIV-11613 H3N2 (black, left), rCIV-23 H3N8 (red, middle) and WY03 H3N2 (blue, right) was examined in triplicate by (A) ELISA, (B) HAI and (C) ELLA assays using 153 human sera samples collected from healthy subjects born between 1934 and 2012 and grouped in 10 years intervals. Dotted black lines indicate the limit of detection of each of the assays. Undetectable titers were assigned a value of 10, 5 and 8 for ELISA, HAI and NAI respectively. Each dot represents the mean titer (represented as log 2 ) of a specific human subject.

Article Snippet: After incubating in block solution (2.5% BSA in PBS) for 1 hour at room temperature, cells were incubated with the following mAbs (1μg/ml) or pAbs (dilution 1:1000) diluted in blocking solution for 1 hour at 37°C: anti-HA A/duck/Shantou/1283/2001 H3N8 goat pAb (BEI Resources NR-34586), A/equine/Miami/1/1963 H3N8 mouse mAb equine 7.1, or A/Hiroshima/52/2005 mouse pAb (BEI Resources F-287) for the staining of rCIV H3N2-, rCIV H3N8- or r/Wy H3N2-infected cells, respectively; anti-NA A/Singapore/1/1957 H2N2 goat pAb (BEI Resources NR-3137), A/equine/Miami/1/1963 H3N8 goat pAb (BEI Resources NR-3145) or hmAb 109219 (kindly provided by James J. Kobie) for the staining of rCIV H3N2-, rCIV H3N8- or r/Wy H3N2-infected cells, respectively; anti-NS1 1A7and 4D4 [ ] mouse mAbs cocktail (1:1, 2 μg/ml).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: PLoS Pathogens

Article Title: Characterizing Emerging Canine H3 Influenza Viruses

doi: 10.1371/journal.ppat.1008409

Figure Lengend Snippet: AIR images of arrays exposed to (A) rCIV-11613 H3N2, (B) rCIV-23 H3N8, (C) HK68 and (D) WI05 viruses. (E) Hierarchical cluster map of the hmAb responses. Values in this map are scaled relative to the antibody producing the strongest response for each virus. Arrows show the antibody clustering groups.

Article Snippet: After incubating in block solution (2.5% BSA in PBS) for 1 hour at room temperature, cells were incubated with the following mAbs (1μg/ml) or pAbs (dilution 1:1000) diluted in blocking solution for 1 hour at 37°C: anti-HA A/duck/Shantou/1283/2001 H3N8 goat pAb (BEI Resources NR-34586), A/equine/Miami/1/1963 H3N8 mouse mAb equine 7.1, or A/Hiroshima/52/2005 mouse pAb (BEI Resources F-287) for the staining of rCIV H3N2-, rCIV H3N8- or r/Wy H3N2-infected cells, respectively; anti-NA A/Singapore/1/1957 H2N2 goat pAb (BEI Resources NR-3137), A/equine/Miami/1/1963 H3N8 goat pAb (BEI Resources NR-3145) or hmAb 109219 (kindly provided by James J. Kobie) for the staining of rCIV H3N2-, rCIV H3N8- or r/Wy H3N2-infected cells, respectively; anti-NS1 1A7and 4D4 [ ] mouse mAbs cocktail (1:1, 2 μg/ml).

Techniques: Virus

Journal: PLoS Pathogens

Article Title: Characterizing Emerging Canine H3 Influenza Viruses

doi: 10.1371/journal.ppat.1008409

Figure Lengend Snippet: MDCK cells were infected with rCIV-11613, rCIV-23, rWY03 or HK68 viruses at a MOI of 3. At 18 hpi, cells were fixed, permeabilized and incubated with 1 μg/ml of the indicated hmAbs. After incubation with a secondary anti-human FITC-conjugated antibody, fluorescence was imaged under a fluorescent microscope. Fluorescence intensity was measured using the ImageJ 1.51s and data was displayed using a Heatmap visualization method. For each virus, the hmAb providing the highest intensity (017–10116 5B03) was considered as 100% and was used to normalize the percentage of reactivity of the rest of the hmAbs. The percentage of reactivity was used to categorized the hmAbs in 6 groups: Group 1: ≥30% reactivity against H3 CIVs and rWY03 H3N2; Group 2: ≥30% reactivity against H3 CIVs, but <30% for rWY03 H3N2; Group 3: ≥30% reactivity against H3N2 rCIV-11613 CIV but <30% for H3N8 rCIV-23 and rWY03 H3N2; Group 4: ≥30% reactivity against H3N2 rCIV-11613 and rWY03 H3N2 but <30% for H3N8 rCIV-23 CIV; Group 5: ≥30% reactivity against rWY03 H3N2 but <30% for rCIV-23 H3N8 and H3N2 rCIV-11613; and, Group 6: non-reactivity or <30% reactivity against rCIV-23 H3N8 and/or H3N2 rCIV-11613, or rWY03. Representative images of the reactivity of the hmAbs against infected cells are shown in the right. Scale represent % of recognition. Scale bars, 200 𝜇m.

Article Snippet: After incubating in block solution (2.5% BSA in PBS) for 1 hour at room temperature, cells were incubated with the following mAbs (1μg/ml) or pAbs (dilution 1:1000) diluted in blocking solution for 1 hour at 37°C: anti-HA A/duck/Shantou/1283/2001 H3N8 goat pAb (BEI Resources NR-34586), A/equine/Miami/1/1963 H3N8 mouse mAb equine 7.1, or A/Hiroshima/52/2005 mouse pAb (BEI Resources F-287) for the staining of rCIV H3N2-, rCIV H3N8- or r/Wy H3N2-infected cells, respectively; anti-NA A/Singapore/1/1957 H2N2 goat pAb (BEI Resources NR-3137), A/equine/Miami/1/1963 H3N8 goat pAb (BEI Resources NR-3145) or hmAb 109219 (kindly provided by James J. Kobie) for the staining of rCIV H3N2-, rCIV H3N8- or r/Wy H3N2-infected cells, respectively; anti-NS1 1A7and 4D4 [ ] mouse mAbs cocktail (1:1, 2 μg/ml).

Techniques: Infection, Incubation, Fluorescence, Microscopy, Virus

Journal: PLoS Pathogens

Article Title: Characterizing Emerging Canine H3 Influenza Viruses

doi: 10.1371/journal.ppat.1008409

Figure Lengend Snippet: Summary of the characteristics of the hmAbs.

Article Snippet: After incubating in block solution (2.5% BSA in PBS) for 1 hour at room temperature, cells were incubated with the following mAbs (1μg/ml) or pAbs (dilution 1:1000) diluted in blocking solution for 1 hour at 37°C: anti-HA A/duck/Shantou/1283/2001 H3N8 goat pAb (BEI Resources NR-34586), A/equine/Miami/1/1963 H3N8 mouse mAb equine 7.1, or A/Hiroshima/52/2005 mouse pAb (BEI Resources F-287) for the staining of rCIV H3N2-, rCIV H3N8- or r/Wy H3N2-infected cells, respectively; anti-NA A/Singapore/1/1957 H2N2 goat pAb (BEI Resources NR-3137), A/equine/Miami/1/1963 H3N8 goat pAb (BEI Resources NR-3145) or hmAb 109219 (kindly provided by James J. Kobie) for the staining of rCIV H3N2-, rCIV H3N8- or r/Wy H3N2-infected cells, respectively; anti-NS1 1A7and 4D4 [ ] mouse mAbs cocktail (1:1, 2 μg/ml).

Techniques:

Journal: PLoS Pathogens

Article Title: Characterizing Emerging Canine H3 Influenza Viruses

doi: 10.1371/journal.ppat.1008409

Figure Lengend Snippet: MDCK cells were infected with the mCherry-expressing rCIV-23 H3N2 (black line), rCIV-23 H3N8 (red line) or rWY03 H3N2 (blue line) recombinant viruses at 0.005 MOI. After 1-hour adsorption at room temperature, cells were incubated with 2-fold serial dilutions of the indicated hmAbs (starting concentration of 0.2 mg/ml) belonging to the Groups 1 (A), 2 (B), 3 (C), 4 (D) or 6 (E). Group 5 hmAbs (≥30% reactivity against rWY03 H3N2 but <30% for rCIV-23 H3N8 and rCIV-23 H3N2) were not included in the assay. At 60 hpi, fluorescence expression was quantified using a fluorescence plate reader and sigmoidal dose response curves were used to calculate the hmAbs concentration that reduced the virus infectivity by 50% (Neutralization titer 50, NT 50 ). Mock-infected and infected cells in the absence of hmAbs were used as controls to normalize the percentage of inhibition. Data show the mean and +/- SDs of the results determined for triplicate. An inhibition of 50% is indicated as a dotted black line.

Article Snippet: After incubating in block solution (2.5% BSA in PBS) for 1 hour at room temperature, cells were incubated with the following mAbs (1μg/ml) or pAbs (dilution 1:1000) diluted in blocking solution for 1 hour at 37°C: anti-HA A/duck/Shantou/1283/2001 H3N8 goat pAb (BEI Resources NR-34586), A/equine/Miami/1/1963 H3N8 mouse mAb equine 7.1, or A/Hiroshima/52/2005 mouse pAb (BEI Resources F-287) for the staining of rCIV H3N2-, rCIV H3N8- or r/Wy H3N2-infected cells, respectively; anti-NA A/Singapore/1/1957 H2N2 goat pAb (BEI Resources NR-3137), A/equine/Miami/1/1963 H3N8 goat pAb (BEI Resources NR-3145) or hmAb 109219 (kindly provided by James J. Kobie) for the staining of rCIV H3N2-, rCIV H3N8- or r/Wy H3N2-infected cells, respectively; anti-NS1 1A7and 4D4 [ ] mouse mAbs cocktail (1:1, 2 μg/ml).

Techniques: Infection, Expressing, Recombinant, Adsorption, Incubation, Concentration Assay, Fluorescence, Virus, Neutralization, Inhibition

Journal: Journal of Clinical Laboratory Analysis

Article Title: Widespread of non‐typeable Haemophilus influenzae with high genetic diversity after two decades use of Hib vaccine in China

doi: 10.1002/jcla.23145

Figure Lengend Snippet: Carriage rates of Haemophilus influenzae isolates in different age group in two sites

Article Snippet: In the present study, H influenzae isolates collected from Chinese children with respiratory infections was monitored in Beijing and discrepancies in the epidemiology between two areas located far apart in China (Beijing City and Youyang County) were identified.

Techniques:

Journal: Journal of Clinical Laboratory Analysis

Article Title: Widespread of non‐typeable Haemophilus influenzae with high genetic diversity after two decades use of Hib vaccine in China

doi: 10.1002/jcla.23145

Figure Lengend Snippet: The results of β‐lactamase activity and antibiotic susceptibility pattern of H influenzae isolates

Article Snippet: In the present study, H influenzae isolates collected from Chinese children with respiratory infections was monitored in Beijing and discrepancies in the epidemiology between two areas located far apart in China (Beijing City and Youyang County) were identified.

Techniques: Activity Assay

Journal: Journal of Clinical Laboratory Analysis

Article Title: Widespread of non‐typeable Haemophilus influenzae with high genetic diversity after two decades use of Hib vaccine in China

doi: 10.1002/jcla.23145

Figure Lengend Snippet: Population snapshot of 190 H influenzae strains revealed by eBURST analysis. The lines indicated the presence of single locus variant links in particular sequence types (STs), which were indicated by circles. The size of the circle corresponded to the number of isolates belonging to the ST. The green ST numbers indicated isolates from Youyang Hospital, the black numbers indicated isolates from Beijing Children's Hospital, and the pink numbers indicates isolates from Youyang Hospital and Beijing Children's Hospital

Article Snippet: In the present study, H influenzae isolates collected from Chinese children with respiratory infections was monitored in Beijing and discrepancies in the epidemiology between two areas located far apart in China (Beijing City and Youyang County) were identified.

Techniques: Variant Assay, Sequencing

Journal: Deutsches Ärzteblatt International

Article Title: The Increase in Invasive Bacterial Infections With Respiratory Transmission in Germany, 2022/2023

doi: 10.3238/arztebl.m2023.0261

Figure Lengend Snippet: Invasive isolates (blood culture, sterile aspirate) of S. pneumoniae (n = 12 478), S. pyogenes (n = 6707), H. influenzae (n = 2037), and N. meningitidis (n = 332) per quarter (sampling date), 2017–2023, Germany (ARS, as of 11 April 2023)

Article Snippet: The Institute for Medical Microbiology at the University Hospital, RWTH Aachen University (appointed as National Reference Center [NRC] for streptococci up to the end of 2022) and the NRC for meningococci and H. influenzae receive voluntarily submitted invasive isolates of H. influenzae, N. meningitidis, S. pneumoniae , and S. pyogenes from all routine laboratories in Germany.

Techniques: Sterility, Sampling

Journal: Deutsches Ärzteblatt International

Article Title: The Increase in Invasive Bacterial Infections With Respiratory Transmission in Germany, 2022/2023

doi: 10.3238/arztebl.m2023.0261

Figure Lengend Snippet: Numbers of invasive isolates (blood culture, sterile aspirate) of S. pneumoniae, S. pyogenes, H. influenzae, and N. meningitidis relative to mean seasonal peak values by quarter, 2017–2019*

Article Snippet: The Institute for Medical Microbiology at the University Hospital, RWTH Aachen University (appointed as National Reference Center [NRC] for streptococci up to the end of 2022) and the NRC for meningococci and H. influenzae receive voluntarily submitted invasive isolates of H. influenzae, N. meningitidis, S. pneumoniae , and S. pyogenes from all routine laboratories in Germany.

Techniques: Sterility

Journal: Deutsches Ärzteblatt International

Article Title: The Increase in Invasive Bacterial Infections With Respiratory Transmission in Germany, 2022/2023

doi: 10.3238/arztebl.m2023.0261

Figure Lengend Snippet: Invasive S. pyogenes isolates (blood culture and aspirate, n = 6707) by age group per 100 000 population, per quarter (sampling date), 2017–2023, Germany (ASR, as of 11 April 2023)

Article Snippet: The Institute for Medical Microbiology at the University Hospital, RWTH Aachen University (appointed as National Reference Center [NRC] for streptococci up to the end of 2022) and the NRC for meningococci and H. influenzae receive voluntarily submitted invasive isolates of H. influenzae, N. meningitidis, S. pneumoniae , and S. pyogenes from all routine laboratories in Germany.

Techniques: Sampling