Comparison of replication competence between two highly pathogenic avian influenza viruses H5N1 and H5N8 in vitro

M. Samy, M. Said, A. H. Mohamed, A. Halim, Mohamed A. Soliman, Moataz Mohamed
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Abstract

ixed infections with respiratory viruses have become frequent at poultry farm business in Egypt. The pandemic of highly pathogenic avian influenza virus (HPAIV) H5N1 (clade 2.2.1.2) has been spreading through the country since 2006. In 2016, the Spread of HPAIV H5N8 (clade 2.3.4.4b) to Egypt via wild birds into commercial poultry flocks made overburden on the economic situation of poultry business. The surveillance efforts conducted by the governmental veterinary authority GOVS showed dominating infection of H5N8 with rare spread of H5N1 in poultry market sector. Thus, the current study try to figure out whether a replication competence exhibited between the two highly pathogenic avian influenza viruses HPAIV H5N1 and H5N8 through designation an in-vitro induced infection of H5N8 and H5N1 in specific pathogen free embryonated chicken eggs (SPF-ECE). A concentration of 10 3 EID50 of HPAIV H5N1 and H5N8 were either sequentially or simultaneously inoculated into the allantoic cavity of SPF-ECE at 48 h of incubation, followed by the collection of allantoic fluid. A quantitative reverse transcription real-time polymerase chain reaction was used to determine the quantitative replication of viral RNA copies of neuraminidase gene of both AIV strains. A replication competence was observed according to which viral strain firstly inoculated. Whereas, H5N1 RNA titers was reduced by ≥4log 10 when H5N8 firstly inoculated. Vice versa H5N8 RNA titers were reduced by ≥4log 10 when H5N1 firstly inoculated. The interference
H5N1和H5N8高致病性禽流感病毒体外复制能力的比较
呼吸道病毒的固定感染在埃及的家禽养殖场已经变得很常见。高致病性禽流感病毒(HPAIV) H5N1(进化支2.2.1.2)大流行自2006年以来一直在全国蔓延。2016年,HPAIV H5N8(分支2.3.4.4b)通过野生鸟类传播到埃及的商业禽群,给家禽业的经济状况带来了沉重的负担。政府兽医当局GOVS开展的监测工作显示,在家禽市场部门H5N8感染占主导地位,H5N1罕见传播。因此,本研究试图通过在特定的无病原体胚性鸡蛋(SPF-ECE)中体外诱导感染H5N8和H5N1两种高致病性禽流感病毒HPAIV H5N1和H5N8之间是否表现出复制能力。将浓度为103eid50的HPAIV H5N1和H5N8依次或同时接种于SPF-ECE的尿囊腔,孵育48 h后收集尿囊液。采用定量反转录实时聚合酶链反应测定两株AIV病毒神经氨酸酶基因RNA拷贝数的定量复制。根据先接种哪一株病毒观察其复制能力。而初次接种H5N8时,H5N1 RNA滴度降低≥4log10。反之,初次接种H5N1时H5N8 RNA滴度降低≥4log 10。的干扰
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