A sandwich ELISA for detecting the hemagglutinin of avian influenza A (H10N8) virus

Abstract

Novel influenza A virus (H10N8) infected human with fatality in China during 2013‐2014. It is important to detect such nonprevalent subtype influenza A virus in clinic and regular surveillance in the early stage for effective control and prevention from the potential pandemic. Unavailability of convenient rapid diagnosis for this subtype virus in resources‐limited setting is an obstacle for timely recognizing human case. In the present study, a panel of mouse H10 specific monoclonal antibodies (mAbs) was generated, two of which were used to develop a sandwich enzyme‐linked immunosorbent assay (ELISA) for detecting the hemagglutinin of avian influenza A (H10N8) virus. ELISA results showed high sensitivity with the lowest detection limit of 0.5HAU/50 μL for live virus, which laid a foundation for clinic use as a promising diagnostic methodology.

Antigen-capture ELISA and immunochromatographic test strip to detect the H9N2 subtype avian influenza virus rapidly based on monoclonal antibodies

“…The procedure for AC-ELISA (Fig. 1 ) was described previously [ 35 , 48 ]. In brief, based on the results of MAb affinity measurements, MAb 3G4 was selected for capture and used to coat a 96-well ELISA plate at 80 ng/well in 100 μl of coating buffer at 4 °C.…”

Development of a monoclonal antibody‐based antigen capture enzyme‐linked immunosorbent assay for detection of H7N9 subtype avian influenza virus

“…Mouse myeloma SP2/0 cells were grown in Dulbecco's modified Eagle's medium (DMEM; Gibco) supplemented with 10% fetal bovine serum (FBS; Gibco) and antibiotics solution consisting of 10 000 units/mL penicillin and 10 000 μg/mL streptomycin (Pen Strep; Gibco). Hybridomas were maintained in DMEM supplemented with 10% FBS, 1% Pen Strep, and 1% hypoxanthine thymidine (HT; Invitrogen) 8 …”

“…Sensitivity was determined using a serial dilution of purified H7-HA protein from A/Zhejiang/DTID-ZJU01/2013(ZJU01) and a standard curve for quantifying H7-HA protein as described previously. 8 We also used serial dilutions of H7 AIVs, including H7N3 (A/duck/ Zhejiang/DK16/2013, DK16), H7N7 (A/chicken/Jiangxi/C25/2014, C25), LP-H7N9 (A/Zhejiang/DTID-ZJU01/2013, ZJU01) and HP-H7N9 (A/Guangdong/HP001/2017, HP001). The detection limit of AC-ELISA was determined based on the highest dilution detected above the cutoff value.…”

“…To this end, an antigen‐capture enzyme‐linked immunosorbent assay (AC‐ELISA) was established based on two specific monoclonal antibodies (mAbs) for detecting viral antigen, resulting in a convenient method for testing many samples simultaneously 7 . Additionally, AC‐ELISA can amplify the response signal through enzyme catalysis, resulting in high sensitivity and specificity 8 . Therefore, an AC‐ELISA was developed in the present study to detect H7N9 subtype AIV rapidly and specifically.…”

Development of an antigen-ELISA and a colloidal gold–based immunochromatographic strip based on monoclonal antibodies for detection of avian influenza A(H5) viruses

“…1) was performed as described previously. 4,16 Monoclonal antibody 4E2 was selected and used to coat a 96-well plate at 160 ng/well in 100 µL of coating buffer. Monoclonal antibody 2F11 was picked for detection and labeled with horseradish peroxidase (HRP; Innoreagents).…”