Development of an Immunochromatographic Strip for Rapid Detection of H7 Subtype Avian Influenza Viruses

Abstract Background H7N9 avian influenza virus (AIV) including highly and low pathogenic viruses have been detected in China since 2013. H7N9 AIV has a high mortality rate after infection in humans, and most human cases have close contacted with poultry in the live poultry market. Therefore, it is necessary to develop a rapid point-of-care testing (POCT) technique for H7N9 AIV detection. Methods The H7N9 AIV was inactivated and purified, and was used as the antigen to immunize BALB/c. Twelve H7-HA specific monoclonal antibodies (McAbs) were produced through the hybridoma technique. The McAb 10A8 was conjugated with colloid gold as detecting antibody; McAb 9B6 was dispensed on the nitrocellulose membran as the capture test line and the Goat-anti mouse IgG antibody was dispensed as control line respectively. The immunochromatographic strip was prepared. Results The analysis of ELISA and virus neutralization test showed that the obtained McAbs specifically recognized H7 HA. Based on the prepared strip, the detection of H7 AIV was achieved within 10 min. No cross-reaction occurred between H7 AIVs and other tested viruses. The detection limit of the strip for H7 was 2.4 log10EID50/0.1 mL for chicken swab samples. Conclusion The McAbs were specific for H7 and the immunochromatographic strip developed in this study was convenient, rapid and reliable for the detection of H7 AIV. The strip could provide an effective method for the rapid and early detection of H7 AIV.

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Development of an Immunochromatographic Strip for Rapid Detection of H7 Subtype Avian Influenza Viruses

10.21203/rs.3.rs-110188/v1 ◽

2020 ◽

Author(s):

Ge Li ◽

Xun Wang ◽

Qingmei Li ◽

Jifei Yang ◽

Xiao Liu ◽

...

Keyword(s):

Avian Influenza ◽

Point Of Care ◽

Influenza Viruses ◽

Control Line ◽

Avian Influenza Viruses ◽

Igg Antibody ◽

Immunochromatographic Strip ◽

Live Poultry ◽

Pathogenic Viruses ◽

H7 Subtype

Abstract Background: H7N9 avian influenza virus (AIV) including highly and low pathogenic viruses have been detected in China since 2013. H7N9 AIV has a high mortality rate after infection in humans, and most human cases have close contacted with poultry in the live poultry market. Therefore, it is necessary to develop a rapid point-of-care testing (POCT) technique for H7N9 AIV detection.Methods: The H7N9 AIV was inactivated and purified, and was used as the antigen to immunize BALB/c. Twelve H7-HA specific monoclonal antibodies (McAbs) were produced through the hybridoma technique. The McAb 10A8 was conjugated with colloid gold as detecting antibody; McAb 9B6 was dispensed on the nitrocellulose membran as the capture test line and the Goat-anti mouse IgG antibody was dispensed as control line respectively. The immunochromatographic strip was prepared.Results: The analysis of ELISA and virus neutralization test showed that the obtained McAbs specifically recognized H7 HA. Based on the prepared strip, the detection of H7 AIV was achieved within 10 min. No cross-reaction occurred between H7 AIVs and other tested viruses. The detection limit of the strip for H7 was 2.4 log10EID50/0.1 mL for chicken swab samples.Conclusion: The McAbs were specific for H7 and the immunochromatographic strip developed in this study was convenient, rapid and reliable for the detection of H7 AIV. The strip could provide an effective method for the rapid and early detection of H7 AIV.

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Development of an Immunochromatographic Strip for Rapid Detection of H9 Subtype Avian Influenza Viruses

Clinical and Vaccine Immunology ◽

10.1128/cvi.00273-07 ◽

2008 ◽

Vol 15 (3) ◽

pp. 569-574 ◽

Cited By ~ 44

Author(s):

Fuhu Peng ◽

Zheng Wang ◽

Shuhui Zhang ◽

Renwei Wu ◽

Sishun Hu ◽

...

Keyword(s):

Avian Influenza ◽

Rapid Detection ◽

Clinical Symptoms ◽

High Specificity ◽

Cloacal Swab ◽

Influenza Viruses ◽

Control Line ◽

Avian Influenza Viruses ◽

Immunochromatographic Strip

ABSTRACT An immunochromatographic strip was developed for the detection of the H9 subtype of avian influenza viruses (H9AIVs) in poultry, using two monoclonal antibodies (MAb), 4C4 for H9AIV hemagglutinin (HA) and 4D4 for nucleoprotein. The 4C4 MAb was labeled with colloidal gold as the detection reagent, and the 4D4 MAb was blotted on the test line while a goat anti-mouse antibody was used on the control line of the nitrocellulose membrane. In comparison with the HA and HA inhibition (HI) tests, the strip was specific for the detection of H9AIV, with a sensitivity at 0.25 HA units within 10 min. Storage of the strips at room temperature for 6 months or at 4°C for 12 months did not change their sensitivity and specificity. Evaluation of the strip with experimental tracheal and cloacal swab samples collected from H9N2-infected chickens revealed that the strip detected the H9N2 viruses on day 3 postinoculation, earlier than the appearance of clinical symptoms. Application of the strip for the analysis of 157 tracheal or cloacal samples from potentially infected chickens on five poultry farms showed that four farms had chickens that were infected with H9AIV. Further characterization of 10 positive and 30 negative randomly selected samples showed that no single sample was false positive or negative, as determined by the standard virus isolation and HI assays. Therefore, the immunochromatographic strip for the detection of H9AIVs has high specificity, sensitivity, and stability. This finding, together with the advantages of rapid detection and easy operation and without the requirement for special skills and equipment, makes the strip suitable for onsite detection and the differentiation of H9AIVs from other viruses in poultry.

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A proof-of-principle study to identify suitable vaccine seed candidates to combat introductions of Eurasian lineage H5 and H7 subtype avian influenza viruses

Avian Pathology ◽

10.1080/03079457.2010.513376 ◽

2010 ◽

Vol 39 (5) ◽

pp. 375-382 ◽

Cited By ~ 7

Author(s):

Maria Serena Beato ◽

Isabella Monne ◽

Marzia Mancin ◽

Elena Bertoli ◽

Ilaria Capua

Keyword(s):

Avian Influenza ◽

Influenza Viruses ◽

Avian Influenza Viruses ◽

Eurasian Lineage ◽

H7 Subtype ◽

Proof Of Principle

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Ecology of H3 avian influenza viruses in Korea and assessment of their pathogenic potentials

Journal of General Virology ◽

10.1099/vir.0.83462-0 ◽

2008 ◽

Vol 89 (4) ◽

pp. 949-957 ◽

Cited By ~ 34

Author(s):

Min-Suk Song ◽

Taek-Kyu Oh ◽

Ho Jin Moon ◽

Dai-Woon Yoo ◽

Eun Ho Lee ◽

...

Keyword(s):

Avian Influenza ◽

Wild Bird ◽

Migratory Birds ◽

Influenza Viruses ◽

Interspecies Transmission ◽

Avian Influenza Viruses ◽

Backyard Poultry ◽

Live Poultry ◽

Live Poultry Markets

To determine the genetic origins of novel H3 avian influenza viruses of chickens and ducks in Korea, genetic characterization of H3 avian influenza viruses isolated from live poultry markets and migratory aquatic birds in South Korea during 2004–2006 was conducted. Phylogenetic analysis revealed that at least four novel genotypes of H3N2 and two genotypes of H3N6 avian influenza viruses were co-circulating in backyard poultry of Korea. The viruses were reassortants between H9N2 viruses of Korean chickens and unknown influenza viruses of migratory birds. Genetic comparison of H3 viruses from live bird markets with those from wild bird isolates revealed that certain gene segments of wild bird isolates are related closely to those of Korean group H9N2 viruses isolated from live poultry markets in 2003. Furthermore, animal-challenge studies demonstrated that the pathogenicity of certain avian H3 influenza viruses was altered due to reassortment, leading to H3 avian influenza viruses in Korea that can potentially expand their host range to include mammals. These studies emphasize the continuing need to monitor backyard poultry at live poultry markets to better understand interspecies transmission and the emergence of novel influenza viruses that have the potential to infect humans.

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Vaccination with Consensus H7 Elicits Broadly Reactive and Protective Antibodies against Eurasian and North American Lineage H7 Viruses

Vaccines ◽

10.3390/vaccines8010143 ◽

2020 ◽

Vol 8 (1) ◽

pp. 143

Author(s):

Gendeal M. Fadlallah ◽

Fuying Ma ◽

Zherui Zhang ◽

Mengchan Hao ◽

Juefu Hu ◽

...

Keyword(s):

Avian Influenza ◽

North American ◽

Human Infection ◽

Influenza Viruses ◽

Avian Influenza Viruses ◽

Lethal Challenge ◽

Protective Antibodies ◽

H7 Subtype ◽

H7n3 Virus

H7 subtype avian influenza viruses have caused outbreaks in poultry, and even human infection, for decades in both Eurasia and North America. Although effective vaccines offer the best protection against avian influenza viruses, antigenically distinct Eurasian and North American lineage subtype H7 viruses require the development of cross-protective vaccine candidates. In this study, a methodology called computationally optimized broadly reactive antigen (COBRA) was used to develop four consensus H7 antigens (CH7-22, CH7-24, CH7-26, and CH7-28). In vitro experiments confirmed the binding of monoclonal antibodies to the head and stem domains of cell surface-expressed consensus HAs, indicating display of their antigenicity. Immunization with DNA vaccines encoding the four antigens was evaluated in a mouse model. Broadly reactive antibodies against H7 viruses from Eurasian and North American lineages were elicited and detected by binding, inhibition, and neutralizing analyses. Further infection with Eurasian H7N9 and North American H7N3 virus strains confirmed that CH7-22 and CH7-24 conferred the most effective protection against hetero-lethal challenge. Our data showed that the consensus H7 vaccines elicit a broadly reactive, protective response against Eurasian and North American lineage H7 viruses, which are suitable for development against other zoonotic influenza viruses.

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