Development of an antigen-capture enzyme-linked immunosorbent assay and immunochromatographic strip based on monoclonal antibodies for detection of H6 avian influenza viruses

Yang, Fan; Xiao, Yixin; Xu, Lixin; Liu, Fumin; Yao, Hang‐Ping; Wu, Nanping; Wu, Haibo

doi:10.1007/s00705-020-04602-w

Cited by 5 publications

(9 citation statements)

References 37 publications

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“…In the detection of actual poultry samples, the AC-ELISA method achieved slightly higher speci city and sensitivity than the ICT strips [43]. This might be related to the visual identi cation method used in the ICT strip.…”

Section: Discussionmentioning

confidence: 98%

“…The positive monoclonal hybridoma cell line that was obtained after three consecutive limiting dilutions was continuously subcultured and then injected into mice intraperitoneally. To obtain MAbs, a Protein G column (GE Healthcare, Chicago, IL, USA) was used to purify ascites collected from the mice injected with the hybridoma cells [43].…”

Section: Viruses and Cellsmentioning

confidence: 99%

“…Isotyping of the MAbs was performed using a Monoclonal Antibody Isotyping Kit (Bio-Rad, Hercules, CA, USA). The a nities of each MAb were measured using ELISA, as described previously [43]. In brief, the ELISA plate was coated with puri ed H9N2 HA protein (20 ng/well) overnight at 4 °C.…”

Section: Isotype and A Nity Of Mabsmentioning

confidence: 99%

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Antigen-Capture ELISA And Immunochromatographic Test Strip To Detect The H9N2 Subtype Avian Influenza Virus Rapidly Based On Monoclonal Antibodies

Xiao

Yang

Liu

et al. 2021

Preprint

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Background: The H9N2 subtype of avian influenza virus (AIV) has become the most widespread subtype of AIV among birds in Asia, which threatens the poultry industry and human health. Therefore, it is important to establish methods for the rapid diagnosis and continuous surveillance of H9N2 subtype AIV.Methods: In this study, an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) and a colloidal gold immunochromatographic test (ICT) strip using monoclonal antibodies (MAbs) 3G4 and 2G7 were established to detect H9N2 subtype AIV. Results: The AC-ELISA method and ICT strip can detect H9N2 subtype AIV quickly, and do not cross-react with other subtype AIVs or other viruses. The detection limit of AC-ELISA was a hemagglutinin (HA) titer of 4 for H9N2 subtype AIV per 100 μl sample, and the limit of detection of the HA protein of AIV H9N2 was 31.5 ng/ml. The ICT strip detection limit was an HA titer of 4 for H9N2 subtype AIV per 100 μl sample. Moreover, both detection methods exhibited good reproducibility and repeatability, with coefficients of variation < 5%. For detection in 200 actual poultry samples, the sensitivities and specificities of AC-ELISA were determined as 93.2% and 98.1%, respectively. The sensitivities and specificities of the ICT strips were determined as 90.9% and 97.4%, respectively. Conclusions: The developed AC-ELISA and ICT strips displayed high specificity, sensitivity, and stability, making them suitable for rapid diagnosis and field investigation of H9N2 subtype AIV.

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Section: Discussionmentioning

confidence: 98%

Section: Viruses and Cellsmentioning

confidence: 99%

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Antigen-Capture ELISA And Immunochromatographic Test Strip To Detect The H9N2 Subtype Avian Influenza Virus Rapidly Based On Monoclonal Antibodies

Xiao

Yang

Liu

et al. 2021

Preprint

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“…Mice were boosted with the same HA protein 3 d before the fusion. 19 Hybridoma culture supernatants were screened by ELISA. Positive monoclonal hybridoma cells were obtained after 3 limiting dilutions and then injected intraperitoneally into mice.…”

mentioning

confidence: 99%

“…The affinities of mAbs were evaluated by ELISA. 19 Briefly, a 96-well plate was coated with purified HA protein (20 ng per well). Monoclonal antibodies at a starting concentration of 1 mg/mL were 2-fold serially diluted and incubated for 1 h. Goat anti-mouse IgG (Novus) diluted to 100 ng/mL was added to the plate and incubated for 30 min.…”

mentioning

confidence: 99%

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

et al. 2021

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Avian influenza A(H5) viruses (avian IAVs) pose a major threat to the economy and public health. We developed an antigen-ELISA (ag-ELISA) and a colloidal gold–based immunochromatographic strip for the rapid detection of avian A(H5) viruses. Both detection methods displayed no cross-reactivity with other viruses (e.g., other avian IAVs, infectious bursal disease virus, Newcastle disease virus, infectious bronchitis virus, avian paramyxovirus). The ag-ELISA was sensitive down to 0.5 hemagglutinin (HA) units/100 µL of avian A(H5) viruses and 7.5 ng/mL of purified H5 HA proteins. The immunochromatographic strip was sensitive down to 1 HA unit/100 µL of avian A(H5) viruses. Both detection methods exhibited good reproducibility with CVs < 10%. For 200 random poultry samples, the sensitivity and specificity of the ag-ELISA were 92.6% and 98.8%, respectively, and for test strips were 88.9% and 98.3%, respectively. Both detection methods displayed high specificity, sensitivity, and stability, making them suitable for rapid detection and field investigation of avian A(H5) viruses.

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Evaluation of panel of neutralising murine monoclonal antibodies and a humanised bispecific antibody against influenza A(H1N1)pdm09 virus infection in a mouse model

et al. 2022

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Development of an antigen-capture enzyme-linked immunosorbent assay and immunochromatographic strip based on monoclonal antibodies for detection of H6 avian influenza viruses

Cited by 5 publications

References 37 publications

Antigen-Capture ELISA And Immunochromatographic Test Strip To Detect The H9N2 Subtype Avian Influenza Virus Rapidly Based On Monoclonal Antibodies

Antigen-Capture ELISA And Immunochromatographic Test Strip To Detect The H9N2 Subtype Avian Influenza Virus Rapidly Based On Monoclonal Antibodies

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

Evaluation of panel of neutralising murine monoclonal antibodies and a humanised bispecific antibody against influenza A(H1N1)pdm09 virus infection in a mouse model

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