A NanoLuc Luciferase Reporter Pseudorabies Virus for Live Imaging and Quantification of Viral Infection.
A NanoLuc Luciferase Reporter Pseudorabies Virus for Live Imaging and Quantification of Viral Infection.
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NanoLuc 荧光素酶报告基因伪狂犬病病毒用于病毒感染的实时成像和定量
DOI：
10.3389/fvets.2020.566446
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发表时间：
2020
影响因子：
3.2
通讯作者：
Sun Y
中科院分区：
文献类型：
--
作者：
Wang Y;Wu H;Wang B;Qi H;Jin Z;Qiu HJ;Sun Y
猪伪狂犬病（Pseudorabies，PR）又称Aujeszky病，是一种急性传染病，给许多国家的养猪业造成重大经济损失。自2011年以来，中国许多接种Bartha-K61疫苗的猪场发生了PR疫情。伪狂犬病病毒（PRV）变异株对猪和小鼠的致病性比以前分离的毒株高。在此，构建了稳定表达NanoLuc（NLuc）荧光素酶与红色荧光蛋白（DsRed）融合的重组PRV（rPRVTJ-NLuc），以追踪病毒在小鼠中的复制和传播。此外，DsRed和NLuc酶在感染的细胞中稳定表达，并且野生型和重组病毒在生长动力学和致病性方面没有显著差异。用103 50%组织培养感染剂量rPRVTJ-NLuc感染7周龄BALB/c小鼠，并进行每日成像。感染rPRVTJ-NLuc的小鼠在感染后4天（dpi）开始显示出稳健的生物发光，生物发光信号随时间增加，在5 dpi达到峰值，至少6 dpi仍可检测到，并在7 dpi消失，同时，增加的通量伴随着病毒从注射部位扩散到上级呼吸道。然而，在脊髓、三叉神经节和来自分离组织的部分脑区域中也观察到信号，而不是在活小鼠中。我们的研究结果描绘了一种新的方法，以快速访问的复制和致病性的新兴PRV在小鼠中。
Pseudorabies (PR), also known as Aujeszky's disease, is an acute infectious disease of pigs, resulting in significant economic losses to the pig industry in many countries. Since 2011, PR outbreaks have occurred in many Bartha-K61-vaccinated pig farms in China. The emerging pseudorabies virus (PRV) variants possess higher pathogenicity in pigs and mice than the strains isolated before. Here, a recombinant PRV (rPRVTJ-NLuc) stably expressing the NanoLuc (NLuc) luciferase fusion with the red fluorescent protein (DsRed) was constructed to trace viral replication and spread in mice. Moreover, both DsRed and NLuc luciferases were stably expressed in the infected cells, and there was no significant difference between wild-type and recombinant viruses in both growth kinetics and pathogenicity. Seven-week-old BALB/c mice were infected with 103 50% tissue culture infective dose rPRVTJ-NLuc and subjected to daily imaging. The mice infected with rPRVTJ-NLuc displayed robust bioluminescence that started 4 days postinfection (dpi), bioluminescence signal increased over time, peaked at 5 dpi, remained detectable for at least 6 dpi, and disappeared at 7 dpi, meanwhile, the increased flux accompanied by the spread of the virus from the injection site to the superior respiratory tract. However, the signal was also observed in the spinal cord, trigeminal ganglion, and partial region of the brain from separated tissues, not in living mice. Our results depicted a new approach to rapidly access the replication and pathogenicity of emerging PRVs in mice.
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影响因子：
7.6
作者：
Hoenen, Thomas;Groseth, Allison;Callison, Julie;Takada, Ayato;Feldmann, Heinz
通讯作者：
Feldmann, Heinz
影响因子：
16.6
作者：
Karlsson, Erik A.;Meliopoulos, Victoria A.;Savage, Chandra;Livingston, Brandi;Mehle, Andrew;Schultz-Cherry, Stacey
通讯作者：
Schultz-Cherry, Stacey
影响因子：
6.7
作者：
Burke CW;Mason JN;Surman SL;Jones BG;Dalloneau E;Hurwitz JL;Russell CJ
通讯作者：
Russell CJ
可视化细胞和活小鼠中呼吸道合胞病毒的复制。
影响因子：
16.6
作者：
Rameix-Welti MA;Le Goffic R;Hervé PL;Sourimant J;Rémot A;Riffault S;Yu Q;Galloux M;Gault E;Eléouët JF
通讯作者：
Eléouët JF
DOI：
10.3390/v8040090
发表时间：
2016-03-29
期刊：
Viruses
影响因子：
--
作者：
Tang YD;Liu JT;Fang QQ;Wang TY;Sun MX;An TQ;Tian ZJ;Cai XH
通讯作者：
Cai XH