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首页> 外文期刊>Avian Diseases >New Approach to Delist Highly Pathogenic Avian Influenza Viruses from BSL3+Select Agents to BSL2 Non-Select Status for Diagnostics and Vaccines
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New Approach to Delist Highly Pathogenic Avian Influenza Viruses from BSL3+Select Agents to BSL2 Non-Select Status for Diagnostics and Vaccines

机译：将高致病性禽流感病毒从BSL3 +选择剂中剔除为BSL2非选择状态的新方法，以用于诊断和疫苗

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摘要

Highly pathogenic avian influenza viruses (AIVs) arc Select Agents in the United States and are required to be handled in bio-containment level-3 enhanced (BSE3+) facilities. Using a reverse genetics system, we attenuated a highly pathogenic virus, with the goal of making it low pathogenic and having it delisted as a Select Agent so that it could be handled in a bio-containment level-2 facility for diagnostic or vaccine production applications. We utilized two approaches to attenuate the target AIV by mutating the highly pathogenic hemagglutinin (HA) cleavage site to be low pathogenic and by replacing the full-length NS gene segment with a naturally truncated 124 amino acid NSI coding gene from A/turkey/Oregon/73 (H7N3) virus (tkOR71 trNS1). To delist an AIV so that it can be handled in a BSL2 facility, the amino acid sequence of the HA cleavage site of the rescued virus must be confirmed to be compatible with a low-pathogenic AIV; it should not plaque in cell culture without supplementation of exogenous trypsin; and intravenous pathotyping in 4-6-wk-old specific-pathogen-free chickens must confirm that the virus is low pathogenic. The candidate A/duck/Vietnana/Baclieu/09/07 (rH5N1/PR8/trNS1) virus with five PR8 internal genes, tkOR71 trNS1 gene, and A/chicken/Indonesia/7/03 NI neuraminidase gene was constructed. The virus was shown to not plaque in cell culture without addition of trypsin. The virus was low pathogenic in the standard intravenous pathotyping rest (IVPI = 0) and also caused no disease in a separate intranasal inoculation test in 4-wk-old specific-pathogen-free chickens, thus demonstrating that the virus is storable for deselection.
机译：高致病性禽流感病毒（AIV）是美国的精选制剂，必须在3级生物安全防护（BSE3 +）设施中处理。使用反向遗传学系统，我们减弱了高致病性病毒，目的是使其具有低致病性并将其从选择试剂中除名，以便可以在生物污染级别2的设施中进行处理以用于诊断或疫苗生产应用。我们利用两种方法通过将高致病性血凝素（HA）切割位点突变为低致病性并通过用来自A /火鸡/俄勒冈州的天然截短的124个氨基酸的NSI编码基因替换全长NS基因片段来减弱目标AIV / 73（H7N3）病毒（tkOR71 trNS1）。为了将AIV除菌，使其可以在BSL2设施中处理，必须确认所拯救病毒的HA切割位点的氨基酸序列与低致病性AIV相容。如果不补充外源胰蛋白酶，则不应在细胞培养中形成斑块。在4-6周龄无特定病原体的鸡中进行静脉病理分型必须确认该病毒是低致病性的。构建了具有五个PR8内部基因，tkOR71 trNS1基因和A /鸡/印度尼西亚/印度尼西亚/ 7/03 NI神经氨酸酶基因的候选A /鸭/越南/ Baclieu / 09/07（rH5N1 / PR8 / trNS1）病毒。在不添加胰蛋白酶的情况下，该病毒在细胞培养中未显示噬菌斑。该病毒在标准的静脉内病理分型休息（IVPI = 0）中是低致病性的，并且在单独的鼻内接种试验中对4周龄无特定病原体的鸡也没有引起疾病，因此证明该病毒可存储以用于选择。

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