Sample
Plasma, tissue homogenates and other biological fluids.
Intended Use
The Influenza A H3N2 (A/Brisbane/10/2007) Hemagglutinin / HA ELISA Pair Set is for the quantitative determination of Influenza A H3N2 (A/Brisbane/10/2007) Hemagglutinin / HA.
This ELISA Pair Set contains the basic components required for the development of sandwich ELISAs.
Contents of Kit
Capture Antibody: 1 mg/mL of rabbit anti-Influenza A H3N2 (A/Brisbane/10/2007) Hemagglutinin / HA monoclonal antibody (in PBS, pH 7.4). Dilute to a working concentration of 2 μg/mL in PBS before coating.
Detection Antibody: 0.2 mg/mL of mouse anti-Influenza A H3N2 (A/Brisbane/10/2007) Hemagglutinin / HA monoclonal antibody conjugated to horseradish-peroxidase (HRP) (in PBS, 50 % HRP-Protector, pH 7.4, store at 4°C). Dilute to working concentration of 0.5 μg/mL in Dilution Buffer before use.
Standard: Each vial contains 1050 ng of recombinant Influenza A H3N2 (A/Brisbane/10/2007) Hemagglutinin / HA. Reconstitute with 1 mL Dilution Buffer. After reconstitution, store at -20°C to -80°C in a manual defrost freezer. A seven-point standard curve using 2-fold serial dilutions in Dilution Buffer, and a high standard of 25000 pg/mL is recommended.
Storage
Capture Antibody: Aliquot and store at -20°C to -80°C for up to 6 months from date of receipt. Avoid repeated freeze-thaw cycles.
Detection Antibody: Store at 4°C and protect it from prolonged exposure to light for up to 6 months from date of receipt. Do not freeze!
Standard: Store lyophilized standard at -20°C to -80°C for up to 6 months from date of receipt. Aliquot and store the reconstituted standard at -80°C for up to 1 month. Avoid repeated freezethaw cycles.
Sensitivity
The minimum detectable dose of Influenza A H3N2 (A/Brisbane/10/2007) Hemagglutinin / HA was determined to be approximately 390.63 pg/ml.
This is defined as at least three times standard deviations above the mean optical density of 10 replicates of the zero standard.
General Description
Influenza (flu) is a respiratory infection in mammals and birds. This virus is divided into three main types (A, B and C). Influenza A is found in a wide variety of bird and mammal species and is further divided into subtypes based on differences in the membrane proteins hemagglutinin (HA) and neuraminidase (NA). H3N2 is a subtype of Influenza A. Hemagglutinin (HA) is a single-pass type I? integral membrane glycoprotein from the influenza virus, and comprises over 80% of the envelope proteins present in the virus particle. The HA is a trimer with a receptor binding pocket on the globular head of each monomer. In natural infection, inactive HA is matured into HA1 and HA2 outside the cell by one or more trypsin-like, arginine-specific endoprotease secreted by the bronchial epithelial cells. Binding of HA to sialic acid- containing receptors on the surface of its target cell brings about the attachment of the virus particle to the cell and forms a endosome. Low pH in endosomes induce an irreversible conformational change in HA2, releasing the hydrophobic portion "fusion peptide". After which, virus penetrates the cell and pours its contents including the RNA genome into the cytoplasm mediated by fusion of the endocytosed virus particle's own membrane and the endosomal membrane. Hemagglutinin plays a major role in the determination of host range restriction and virulence.
Citations
Publication ()
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Seroepidemiological Evidence of Subtype H3N8 Influenza Virus Infection among Pet Dogs in China
Pei Zhou,San Huang,Weijie Zeng,Xin Zhang,Lifang Wang,Xinliang Fu,Shoujun Li
Applications: ELISA
Reactive species: Human
"Abstract: The H3N8 virus and the H3N2 virus are the main subtypes of canine influenza virus (CIV). H3N8 CIV mainly circulates in America, and H3N2 CIV mainly circulates in Asia. However, there was an outbreak of the Asian H3N2 virus in the United States (US) in 2015. Thus, it is important to evaluate the presence of subtype H3N8 virus in dogs in China. From May 2015 to November 2015, 600 sera from pet dogs were collected from Guangzhou, Shanghai, Beijing and Shenzhen for hemagglutination inhibition (HI) assays and microneutralization (MN) assays. Fifty-two (8.66%) of the 600 sera were positive for the subtype H3N2 virus, which matched the previous reports. Five (0.83%) of 600 sera were positive for the subtype H3N8 virus (H3N8 EIV or H3N8 AIV or H3N8 CIV), which is the first report of subtype H3N8 virus infection among dogs in China and remind us to play more attention to this subtype virus. Therefore, further serological and virological surveillance of influenza virus infection among dogs in China is imperative."
Article snippet: Additionally, the specimens with an HI titer > 20 against H3N8 virus were further evaluated for human H3N2 influenza virus (HuIV) antibody using an enzyme-linked immunosorbent assay (ELISA, H3N2 HA1 Hemagglutinin, HA1 ELISA Kit, Prod. No.: DEIA252, Creative Diagnostics company) according to the manufacturer’s instructions. 
Figure 1. HI titers of control sera against reference virus strains.
COA
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