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Influenza A H5N1 (Avian Flu) Hemagglutinin, HA ELISA Kit

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[#DEIA250] Influenza A H5N1 (Avian Flu) Hemagglutinin, HA ELISA Kit

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DEIA250 | Influenza A H5N1 (Avian Flu) Hemagglutinin, HA ELISA Kit, 5 plates
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(1) Review analysis and impact of co-circulating H5N1 and H9N2 avian influenza viruses in Bangladesh.[TOP]

Pubmed ID :29781424
Publication Date : //
Almost the full range of 16 haemagglutinin (HA) and nine neuraminidase subtypes of avian influenza viruses (AIVs) has been detected either in waterfowl, land-based poultry or in the environment in Bangladesh. AIV infections in Bangladesh affected a wide range of host species of terrestrial poultry. The highly pathogenic avian influenza (AI) H5N1 and low pathogenic AI H9N2 were found to co-circulate and be well entrenched in the poultry population, which has caused serious damage to the poultry industry since 2007. By reviewing the available scientific literature, the overall situation of AIVs in Bangladesh is discussed. All Bangladeshi (BD) H5N1 and H9N2 AIV sequences available at GenBank were downloaded along with other representative sequences to analyse the genetic diversity among the circulating AIVs in Bangladesh and to compare with the global situation. Three different H5N1 clades, 2.2.2, 2.3.2.1 and 2.3.4.2, have been detected in Bangladesh. Only 2.3.2.1a is still present. The BD LP H9N2 viruses mostly belonged to the H9 G1 lineage but segregated into many branches, and some of these shared internal genes with HP viruses of subtypes H7N3 and H5N1. However, these reassortment events might have taken place before introduction to Bangladesh. Currently, H9N2 viruses continue to evolve their HA cleavage, receptor binding and glycosylation sites. Multiple mutations in the HA gene associated with adaptation to mammalian hosts were also observed. Strict biosecurity at farms and gradual phasing out of live-bird markets could be the key measures to better control AIVs, whereas stamping out is not a practicable option in Bangladesh. Vaccination also could be an additional tool, which however, requires careful planning. Continuous monitoring of AIVs through systematic surveillance and genetic characterisation of the viruses remains a hallmark of AI control.

Authors : Parvin Rokshana, Begum Jahan Ara, Nooruzzaman Mohammed, Chowdhury Emdadul Haque, Islam Mohammad Rafiqul, Vahlenkamp Thomas W,



(2) Genetic analysis and biological characteristics of different internal gene origin H5N6 reassortment avian influenza virus in China in 2016.[TOP]

Pubmed ID :29778197
Publication Date : //
Clade 2.3.4.4 of H5N6 subtype Avian Influenza Viruses (AIVs) has become dominant clade in South-East Asia. So far, a total of 16 cases of human infection, including 6 deaths, have been confirmed since 2014. In this study, we systematically investigated the genetic evolution and biological characteristics of these viruses. We first carried out phylogenetic and statistical analysis of all H5N6 viruses that were downloaded from Influenza Research Database, GISAID and isolates from our lab. We found that H5N6 AIVs continued to reassort with other AIVs subtypes since 2014. Among these H5N6 reassortments, four main gene types were identified: A (internal genes of H5N1-origin), B (PB2 of H6-origin, and others of H5N1-origin), C (internal genes of H9-origin) and D (PB2 of H6-origin and PB1of H3-origin, and others of H5N1). In addition, after several years of evolution, gene type D is currently the dominant gene type. To systematically compare the genetic and evolutionary characteristics and pathogenicity of these viruses, four H5N6 AIVs of different gene types were selected for further analysis. S4, XZ6, GD1602 and YZ587 virus represented gene type A, B, C and D, respectively. Their NA genes were all originated from H6 and their whole genome showed a high similarity with human isolates. All these isolates could both bind with SA-α2,3 Gal and SA-α2,6 Gal receptors. Pathogenicity test showed that these viruses were highly pathogenic in chickens, while YZ587 showed the lowest virulence. Moreover, XZ6 and S4 viruses were highly pathogenic in ducks and moderately pathogenic in mice, while GD1602 and YZ587 viruses were no-pathogenic in these animals. Interestingly, GD1602 and YZ587-like viruses were responsible for 4 and 2 human infection cases in 2016, respectively. Therefore, our study showed that the YZ587 virus which has mixed internal genes, showed lower virulence in avian species and mammals compared to other genotype viruses. Overall, our findings suggest that the H5N6 avian influenza virus is undergoing constantly evolving and reassortment. Thus, our study highlights the necessary of continued surveillance of the H5N6 AIVs in birds and paying close attention to the spread of these novel reassortment viruses.

Authors : Sun Wenqiang, Li Jiaxin, Hu Jiao, Jiang Daxiu, Xing Chaonan, Zhan Tiansong, Liu Xiufan,



(3) Public health response to outbreaks of Avian Influenza A(H5N2) and (H5N1) among poultry - British Columbia, December 2014-February 2015.[TOP]

Pubmed ID :29769935
Publication Date : //
In December 2014, the first detection in Canada of a highly pathogenic avian influenza A (HPAI) virus was reported in poultry within the Fraser Health Authority of British Columbia. It was the second outbreak of HPAI from Eurasian H5 reassortment viruses in North America. The Fraser Health Authority provided the lead public health coordination for this response as well as consultation and support to the occupational health response. The public health response focused on contact tracing, monitoring and follow-up for household, farm worker and other community contacts exposed on the affected farms. A total of 50 contacts were identified. Contacts received daily active monitoring by public health nurses for seven days from their last exposure and were advised to self-monitor until day 10. All contacts and other household members were recommended seasonal influenza vaccination to protect against further possible reassortment with human influenza viruses circulating within the community at the time. A total of 26 (52%) contacts were recommended chemoprophylaxis for ongoing exposure to the affected barns and flocks, of whom only 11 (42%) initiated this. During the seven-day active surveillance period, four contacts developed acute respiratory symptoms and influenza B was identified in one individual. Local area health care providers and acute care facilities were alerted to the outbreak and public messaging was provided regarding the human health risks from avian influenza. Collaboration between health and agriculture at the local, regional, provincial and federal levels was key to a rapid response to this outbreak.

Authors : Murti M, Skowronski D, Lem M, Fung C, Klar S, Bigham M, Loadman S, Chambers C, Pritchard J, Lee V,



(4) Comparative analysis of MicroRNA expression in dog lungs infected with the H3N2 and H5N1 canine influenza viruses.[TOP]

Pubmed ID :29772263
Publication Date : //
MicroRNAs, a class of noncoding RNAs 18 to 23 nucleotides (nt) in length, play critical roles in a wide variety of biological processes. The objective of this study was to examine differences in microRNA expression profiles derived from the lungs of beagle dogs infected with the avian-origin H3N2 canine influenza virus (CIV) or the highly pathogenic avian influenza (HPAI) H5N1 virus (canine-origin isolation strain). After dogs were infected with H3N2 or H5N1, microRNA expression in the lungs was assessed using a deep-sequencing approach. To identify the roles of microRNAs in viral pathogenicity and the host immune response, microRNA target genes were predicted, and their functions were analyzed using bioinformatics software. A total of 229 microRNAs were upregulated in the H5N1 infection group compared with those in the H3N2 infection group, and 166 microRNAs were downregulated. MicroRNA target genes in the H5N1 group were more significantly involved in metabolic pathways, such as glycerolipid metabolism and glycerophospholipid metabolism, than those in the H3N2 group. The inhibition of metabolic pathways may lead to appetite loss, weight loss and weakened immunity. Moreover, miR-485, miR-144, miR-133b, miR-4859-5p, miR-6902-3p, miR-7638, miR-1307-3p and miR-1346 were significantly altered microRNAs that potentially led to the inhibition of innate immune pathways and the heightened pathogenicity of H5N1 compared with that of H3N2 in dogs. This study deepens our understanding of the complex relationships among microRNAs, the influenza virus-mediated immune response and immune injury in dogs.

Authors : Zheng Yun, Fu Xinliang, Wang Lifang, Zhang Wenyan, Zhou Pei, Zhang Xin, Zeng Weijie, Chen Jidang, Cao Zongxi, Jia Kun, Li Shoujun,



(5) Genome-wide profiling of microRNAs reveals novel insights into the interactions between H9N2 avian influenza virus and avian dendritic cells.[TOP]

Pubmed ID :29743596
Publication Date : //
The antigen-presenting ability of dendritic cells (DCs) plays an important and irreplaceable role in recognising and clearing viruses. Antiviral responses must rapidly defend against infection while minimising inflammatory damage, but the mechanisms that regulate the magnitude of response within an infected cell are not well understood. MicroRNAs (microRNAs), small non-coding RNAs, can regulate mouse or avian DCs to inhibit the infection and replication of avian influenza virus (AIV). Here, we performed a global analysis to understand how avian DCs respond to H9N2 AIV and provide a potential mechanism to explain how avian microRNAs can defend against H9N2 AIV replication. First, we found that both active and inactive H9N2 AIV enhanced the ability of DCs to present antigens and activate T lymphocytes. Next, total microarray analyses suggested that H9N2 AIV stimulation involved protein localisation, nucleotide binding, leucocyte transendothelial migration and MAPK signalling. Moreover, we constructed 551 transcription factor (TF)-miRNA-mRNA loops based on the above analyses. Furthermore, we found that the haemagglutinin (HA) fragment, neither H5N1-HA or H9N2-HA, could not activate DCs, while truncated HA greatly increased the immune function of DCs by activating ERK and STAT3 signalling pathways. Lastly, our results not only suggested that gga-miR1644 targets muscleblind-like protein 2 (MBNL2) to enhance the ability of avian DCs to inhibit virus replication, but also suggested that gga-miR6675 targets the nuclear localisation sequence of polymerase basic protein 1 (PB1) to trigger the silencing of PB1 genes, resulting in the inhibition of H9N2 AIV replication. Altogether, our innovative study will shed new light on the role of avian microRNAs in evoking avian DCs and inhibiting virus replication.

Authors : Lin Jian, Xia Jing, Zhang Tian, Zhang Keyun, Yang Qian,



(6) N-myc downstream-regulated gene 1 facilitates influenza A virus replication by suppressing canonical NF-κB signaling.[TOP]

Pubmed ID :29730307
Publication Date : //
The highly pathogenic avian influenza (HPAI) A/H5N1 virus hijacks host cellular machinery to complete its life cycle; identification of the host factors involved in viral replication may facilitate antiviral drug development. Here, we first characterize a metastasis suppressor, N-myc downstream-regulated gene 1 (NDRG1), and showed that it plays a crucial role in H5N1 viral replication. We found that H5N1 infection upregulated NDRG1 mRNA and protein expression. Overexpression of NDRG1 released approximately 4-fold more virions compared to the control group, whereas knockdown of NDRG1 resulted in a drop in viral RNA and protein production. Further investigation revealed that NDRG1 facilitated HPAI A/H5N1 viral replication by suppressing the canonical NF-κB signaling pathway. Furthermore, our results also showed that the NDRG1 mRNA level was mainly stimulated by M1 and PB1 viral proteins. Overall, our results suggest that NDRG1 plays a positive role in HPAI replication by suppressing the canonical NF-κB signaling pathway.

Authors : Chen Lin, Xing Chao, Ma Guoyao, Luo Jing, Su Wen, Li Meng, Shi Qiumei, He Hongxuan,



(7) Development and evaluation of a real-time RT-PCR assay for detection of a novel avian influenza A (H5N6) virus.[TOP]

Pubmed ID :29729298
Publication Date : //
As of Aug 25, 2017, 17 incidences of human infection and 6 deaths due to the novel H5N6 virus have been reported in China. Genetic analysis of the viral genome revealed that this reassortant virus is highly pathogenic to poultry, and that the virus has a risk of transmission to humans. Accordingly, the development of a rapid, sensitive, and specific molecular diagnostic assay is critical for public health. In this study, a real-time reverse-transcription PCR (RT-PCR) assay was developed to specifically detect the novel H5N6 virus, with primer pairs targeting the hemagglutinin and neuraminidase gene sequences of this virus. RNA was extracted from throat swab specimens from patients with influenza-like illness (ILIs), and environmental samples were collected from live poultry markets (LPMs) for H5N6 virus detection by real-time RT-PCR. The method was demonstrated to enable specific detection of the avian H5N6 virus, with no cross-reactivity with seasonal influenza viruses (H1N1, H1N1 pdm09, H3N2 or B); H5N1, H7N9, H9N2 viruses; or other human respiratory viruses. The detection limit of the assay was 1.0 × 10 copies per reaction for N6 and 1.0 × 10 copies per reaction for H5 assays. The assay is a powerful tool for rapid, sensitive, and specific detection of H5N6 virus infection in specimens derived from humans, animals, and the environment.

Authors : Zhang Rusheng, Yao Dong, Chen Jingfang, Ye Wen, Ou Xinhua, Chen Tianmu, Sun Biancheng,



(8) Migratory Whooper Swans Cygnus cygnus Transmit H5N1 Virus between China and Mongolia: Combination Evidence from Satellite Tracking and Phylogenetics Analysis.[TOP]

Pubmed ID :29728621
Publication Date : //
In late 2014, a highly pathogenic avian influenza (hereafter HPAI) H5N1 outbreak infected whooper swans Cygnus cygnus wintering at the Sanmenxia Reservoir area, China, and raised concerns about migratory linkages between wintering and breeding grounds of whooper swans. In this study, 61 swans were satellite tracked from 2013 to 2016 to determine the spatial association of their migration routes and H5N1 outbreaks, and 3596 fecal samples were collected along the migration routes for virology testing. Swans departed the wintering grounds and migrated along the Yellow River, and flew over the Yin Mountains in China. The Brownian bridge movement model showed there was a high degree of spatiotemporal overlap between the core use area along the spring migration pathway and historical H5N1 events in China and Mongolia from 2005 to 2015. The H5N1 strain was isolated and phylogenetic analyses confirmed that the HA gene sequence generated is genetically similar to that of the epidemic strain at a previous wintering site (the Sanmenxia Reservoir area) along its flyway. Our results identified a previously unknown migratory link of whooper swans in central China with Mongolia and confirmed that the swans could carry the HPAI H5N1 virus during migration, resulting in long-distance transmission.

Authors : Li Shuhong, Meng Weiyue, Liu Dongping, Yang Qiqi, Chen Lixia, Dai Qiang, Ma Tian, Gao Ruyi, Ru Wendong, Li Yunfeng, Yu Pengbo, Lu Jun, Zhang Guogang, Tian Huaiyu, Chai Hongliang, Li Yanbing,



(9) Experimental infection of H5N1 and H5N8 highly pathogenic avian influenza viruses in Northern Pintail (Anas acuta).[TOP]

Pubmed ID :29726612
Publication Date : //
The wide geographic spread of Eurasian Goose/Guangdong lineage highly pathogenic avian influenza (HPAI) clade 2.3.4.4 viruses by wild birds is of great concern. In December 2014, an H5N8 HPAI clade 2.3.4.4 Group A (2.3.4.4A) virus was introduced to North America. Long-distance migratory wild aquatic birds between East Asia and North America, such as Northern Pintail (Anas acuta), were strongly suspected of being a source of intercontinental transmission. In this study, we evaluated the pathogenicity, infectivity and transmissibility of an H5N8 HPAI clade 2.3.4.4A virus in Northern Pintails and compared the results to that of an H5N1 HPAI clade 2.3.2.1 virus. All of Northern Pintails infected with either H5N1 or H5N8 virus lacked clinical signs and mortality, but the H5N8 clade 2.3.4.4 virus was more efficient at replicating within and transmitting between Northern Pintails than the H5N1 clade 2.3.2.1 virus. The H5N8-infected birds shed high titre of viruses from oropharynx and cloaca, which in the field supported virus transmission and spread. This study highlights the role of wild waterfowl in the intercontinental spread of some HPAI viruses. Migratory aquatic birds should be carefully monitored for the early detection of H5 clade 2.3.4.4 and other HPAI viruses.

Authors : Kwon J-H, Lee D-H, Swayne D E, Noh J-Y, Yuk S-S, Jeong S, Lee S-H, Woo C, Shin J-H, Song C-S,



(10) Substitution of D701N in the PB2 protein could enhance the viral replication and pathogenicity of Eurasian avian-like H1N1 swine influenza viruses.[TOP]

Pubmed ID :29717109
Publication Date : //
Eurasian avian-like H1N1 (EA H1N1) swine influenza viruses (SIVs) have become predominant in pig populations in China and have recently been reported to have the most potential to raise the next pandemic in humans. The mutation D701N in the PB2 protein, which accounts for 31% of H1N1 SIVs, has previously been shown to contribute to the adaptation of the highly pathogenic H5N1 or H7N7 avian influenza viruses in mammals. However, little is known of the effects of this substitution on the EA H1N1 viruses. Herein, we investigated the contributions of 701N in the PB2 protein to an EA H1N1 SIV (A/Hunan/42443/2015(H1N1), HuN EA-H1N1), which had 701D in the PB2 protein. Our results found that viral polymerase activity, viral replication, and pathogenicity in mice were indeed enhanced due to the introduction of 701N into the PB2 protein, and the increased viral growth was partly mediated by the host factor importin-α7. Thus, substantial attention should be paid to the D701N mutation in pig populations.

Authors : Liu Suli, Zhu Wenfei, Feng Zhaomin, Gao Rongbao, Guo Junfeng, Li Xiyan, Liu Jia, Wang Dayan, Shu Yuelong,