Production and Identification of Monoclonal Antibodies and Development of a Sandwich ELISA for Detection of the H3-subtype Avian Influenza Virus Antigen

Overview

Journal AMB Express

Date 2020 Mar 15

PMID 32170425

Citations 7

Authors

Sisi Luo

Xianwen Deng

Zhixun Xie

Jiaoling Huang

Minxiu Zhang

Meng Li

Liji Xie

Dan Li

Qing Fan

Sheng Wang

TingTing Zeng

Yanfang Zhang

Zhiqin Xie

Affiliations

Soon will be listed here.

Abstract

The H3 subtype of avian influenza virus (AIV) is widespread in avian species and is frequently isolated in surveillance projects; thus, we have developed a more effective diagnostic approach of a monoclonal antibody (mAb)-based sandwich ELISA for the H3 AIV detection. First, we have produced the essential reagent of mAb against AIV H3 strains with the development of an mAb-Mouse immunization with a purified H3-subtype AIV strain and cell fusion to generate hybridoma cells. These cells were screened with hemagglutination inhibition (HI) tests, and optimal cells were subcloned. We chose a hybridoma cell line that steadily secreted a specific H3-subtype AIV mAb, designated 9F12, that belongs to the IgG1 subclass and has a K-type light chain. 9F12 was shown to bind specifically to the H3-subtype AIV antigen by both immunofluorescence assay and Western blot analysis. Finally, a 9F12-based sandwich ELISA was successfully developed and used to specifically test for this antigen. The sandwich ELISA conditions were optimized, and the specificity and sensitivity were validated. The results for clinical sample detection were consistent with viral isolation. Consequently, the 9F12-based sandwich ELISA is a specific, sensitive, robust, rapid and versatile diagnostic tool for H3-subtype AIV and provides a promising strategy for effective influenza virus prevention and control.

Citing Articles

A Rapid Detection Method for H3 Avian Influenza Viruses Based on RT-RAA.

Li J, Cui H, Zhang Y, Wang X, Liu H, Mu Y Animals (Basel). 2024; 14(17).

PMID: 39272386 PMC: 11393923. DOI: 10.3390/ani14172601.

Evaluating the efficiency of ELISA, monoplex and multiplex probe-based real-time reverse-transcription PCR assays in the detection of SARS-CoV-2 (COVID-19) and influenza A and B viruses: A cross-sectional study.

Mosadegh M, Jalili S, Pourmand M, Erfani Y, Panji M Health Sci Rep. 2024; 7(6):e2140.

PMID: 38915351 PMC: 11194474. DOI: 10.1002/hsr2.2140.

Development of a Double-Antibody Sandwich ELISA Based on a Monoclonal Antibody against the Viral NS1 Protein for the Detection of Chicken Parvovirus.

Zhang M, Liao J, Xie Z, Zhang Y, Luo S, Li M Pathogens. 2024; 13(3).

PMID: 38535564 PMC: 10976255. DOI: 10.3390/pathogens13030221.

Editorial: High-impact respiratory RNA virus diseases.

Petrone-Garcia V, Castellanos-Huerta I, Tellez-Isaias G Front Vet Sci. 2023; 10:1273650.

PMID: 37675076 PMC: 10478262. DOI: 10.3389/fvets.2023.1273650.

Monoclonal antibody targeting a novel linear epitope on nucleoprotein confers pan-reactivity to influenza A virus.

Gu M, Jiao J, Liu S, Zhao W, Ge Z, Cai K Appl Microbiol Biotechnol. 2023; 107(7-8):2437-2450.

PMID: 36820898 PMC: 9947902. DOI: 10.1007/s00253-023-12433-3.

References

He W, Li G, Zhu H, Shi W, Wang R, Zhang C . Emergence and adaptation of H3N2 canine influenza virus from avian influenza virus: An overlooked role of dogs in interspecies transmission. Transbound Emerg Dis. 2018; 66(2):842-851. DOI: 10.1111/tbed.13093. View

Li C, Yu M, Liu L, Sun H . Characterization of a novel H3N2 influenza virus isolated from domestic ducks in China. Virus Genes. 2016; 52(4):568-72. DOI: 10.1007/s11262-016-1323-0. View

Peng Y, Xie Z, Liu J, Pang Y, Deng X, Xie Z . Visual detection of H3 subtype avian influenza viruses by reverse transcription loop-mediated isothermal amplification assay. Virol J. 2011; 8:337. PMC: 3154870. DOI: 10.1186/1743-422X-8-337. View

Chen L, Ruan F, Sun Y, Chen H, Liu M, Zhou J . Establishment of sandwich ELISA for detecting the H7 subtype influenza A virus. J Med Virol. 2019; 91(6):1168-1171. DOI: 10.1002/jmv.25408. View

Oshansky C, Wong S, Jeevan T, Smallwood H, Webby R, Shafir S . Seasonal influenza vaccination is the strongest correlate of cross-reactive antibody responses in migratory bird handlers. mBio. 2014; 5(6):e02107. PMC: 4324241. DOI: 10.1128/mBio.02107-14. View

Huang L, Wei Y, Xia D, Liu D, Zhu H, Wu H . A broad spectrum monoclonal antibody against porcine circovirus type 2 for antigen and antibody detection. Appl Microbiol Biotechnol. 2019; 103(8):3453-3464. DOI: 10.1007/s00253-019-09715-0. View

Tong S, Li Y, Rivailler P, Conrardy C, Alvarez Castillo D, Chen L . A distinct lineage of influenza A virus from bats. Proc Natl Acad Sci U S A. 2012; 109(11):4269-74. PMC: 3306675. DOI: 10.1073/pnas.1116200109. View

Darcy E, Leonard P, Fitzgerald J, Danaher M, OKennedy R . Purification of antibodies using affinity chromatography. Methods Mol Biol. 2010; 681:369-82. DOI: 10.1007/978-1-60761-913-0_20. View

Tang Q, Wang J, Bao J, Sun H, Sun Y, Liu J . A multiplex RT-PCR assay for detection and differentiation of avian H3, H5, and H9 subtype influenza viruses and Newcastle disease viruses. J Virol Methods. 2012; 181(2):164-9. DOI: 10.1016/j.jviromet.2012.02.003. View

10.

Shichinohe S, Okamatsu M, Sakoda Y, Kida H . Selection of H3 avian influenza viruses with SAα2,6Gal receptor specificity in pigs. Virology. 2013; 444(1-2):404-8. DOI: 10.1016/j.virol.2013.07.007. View

11.

Wang Z, Yu J, Thomas M, Sreenivasan C, Hause B, Wang D . Pre-exposure with influenza A virus A/WSN/1933(H1N1) resulted in viral shedding reduction from pigs challenged with either swine H1N1 or H3N2 virus. Vet Microbiol. 2018; 228:26-31. PMC: 6314213. DOI: 10.1016/j.vetmic.2018.11.008. View

12.

Sreenivasan C, Jandhyala S, Luo S, Hause B, Thomas M, Knudsen D . Phylogenetic Analysis and Characterization of a Sporadic Isolate of Equine Influenza A H3N8 from an Unvaccinated Horse in 2015. Viruses. 2018; 10(1). PMC: 5795444. DOI: 10.3390/v10010031. View

13.

Solorzano A, Foni E, Cordoba L, Baratelli M, Razzuoli E, Bilato D . Cross-Species Infectivity of H3N8 Influenza Virus in an Experimental Infection in Swine. J Virol. 2015; 89(22):11190-202. PMC: 4645675. DOI: 10.1128/JVI.01509-15. View

14.

Zhang L, Li Z, Jin H, Hu X, Su J . Development and application of a monoclonal antibody-based blocking ELISA for detection of antibodies to Tembusu virus in multiple poultry species. BMC Vet Res. 2018; 14(1):201. PMC: 6019803. DOI: 10.1186/s12917-018-1537-6. View

15.

Tong S, Zhu X, Li Y, Shi M, Zhang J, Bourgeois M . New world bats harbor diverse influenza A viruses. PLoS Pathog. 2013; 9(10):e1003657. PMC: 3794996. DOI: 10.1371/journal.ppat.1003657. View

16.

Teng Q, Shen W, Yan D, Yan L, Li X, Li G . Development of a TaqMan MGB RT-PCR for the rapid detection of H3 subtype avian influenza virus circulating in China. J Virol Methods. 2015; 217:64-9. DOI: 10.1016/j.jviromet.2015.02.025. View

17.

Ito M, Yamayoshi S, Murakami K, Saito K, Motojima A, Nakaishi K . Characterization of Mouse Monoclonal Antibodies Against the HA of A(H7N9) Influenza Virus. Viruses. 2019; 11(2). PMC: 6410113. DOI: 10.3390/v11020149. View

18.

Beuttemmuller E, Woodward A, Rash A, Dos Santos Ferraz L, Alfieri A, Alfieri A . Characterisation of the epidemic strain of H3N8 equine influenza virus responsible for outbreaks in South America in 2012. Virol J. 2016; 13:45. PMC: 4799594. DOI: 10.1186/s12985-016-0503-9. View

19.

Song D, Kang B, Lee C, Jung K, Ha G, Kang D . Transmission of avian influenza virus (H3N2) to dogs. Emerg Infect Dis. 2008; 14(5):741-6. PMC: 2600237. DOI: 10.3201/eid1405.071471. View

20.

Guan L, Shi J, Kong X, Ma S, Zhang Y, Yin X . H3N2 avian influenza viruses detected in live poultry markets in China bind to human-type receptors and transmit in guinea pigs and ferrets. Emerg Microbes Infect. 2019; 8(1):1280-1290. PMC: 6746299. DOI: 10.1080/22221751.2019.1660590. View