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用于开发马 rhinopneumonitis 疫苗的 AK-2011 株。

AK-2011 strain for the development of a vaccine against equine rhinopneumonitis.

机构信息

Laboratory of Cell Cultures and Culture Media, LLP «SPE DiaVak-ABN», Almaty, Kazakhstan.

Virology laboratory, LLP «SPE DiaVak-ABN», Almaty, Kazakhstan.

出版信息

Transbound Emerg Dis. 2022 Sep;69(5):e1972-e1981. doi: 10.1111/tbed.14531. Epub 2022 Apr 5.

Abstract

Equine rhinopneumonitis is an acute, highly contagious disease found virtually worldwide. The purpose of the studies presented in this paper is to develop a technology for the manufacture of a cell-derived equine rhinopneumonitis vaccine, as well as to assess the safety and immunogenicity of the newly developed vaccine in laboratory animals model. The object of the studies was the AK-2011 strain isolated from the horses suffering from rhinopneumonitis during an outbreak of abortions. The viability of the AK-2011 strain was assessed using a continuous line of calf trachea cells, a continuous line of calf kidney cells, a continuous line of sheep kidney cells, a continuous line of bovine kidney cells, a continuous line of green monkey kidney cells, a continuous line of Syrian hamster kidney cells, a primary trypsinized culture of horse kidney cells grown in tubes and flasks and the AK-2011 laboratory strain of equine rhinopneumonitis virus with biological activity of 6.0 lg TCID . Sequencing and polymerase chain reaction analysis were performed. The virus isolated from the ORF68 gene in Kazakhstan appeared to be the most similar to the T-953 and 2222-03 strains isolated in the USA and Australia, respectively, in terms of phylogenetics. As to primary infections, cytopathic effects (CPEs) induced by the AK-2011 virus stain (dilution 10 ) in calf trachea and horse kidney cell cultures were stable from the first to tenth passages, with biological activity of 5.75-6.00 lg TCID . CPEs caused by the virus were apparent on days 2-3, further developed intensively and extended to 60-80% of the cell monolayer on days 5-7. The vaccine results can be used to immunize horses on farms against rhinopneumonia, and horses should be immunized twice with an interval of 2-3 months.

摘要

马传染性鼻肺炎是一种几乎在全世界范围内都存在的急性、高度传染性疾病。本论文中所进行的研究旨在开发一种细胞衍生马传染性鼻肺炎疫苗的制造技术,并评估新开发的疫苗在实验室动物模型中的安全性和免疫原性。研究对象是从流产暴发期间患有鼻肺炎的马匹中分离出的 AK-2011 株。使用连续传代的牛气管细胞系、连续传代的牛肾细胞系、连续传代的绵羊肾细胞系、连续传代的牛肾细胞系、连续传代的绿猴肾细胞系、连续传代的叙利亚仓鼠肾细胞系、在管瓶中生长的原代胰蛋白酶化马肾细胞培养物以及具有 6.0 lgTCID 的 AK-2011 实验室株马传染性鼻肺炎病毒来评估 AK-2011 株的活力。进行了测序和聚合酶链反应分析。从哈萨克斯坦分离出的病毒在 ORF68 基因方面与分别从美国和澳大利亚分离出的 T-953 和 2222-03 株最为相似,在系统发育方面。至于初次感染,AK-2011 病毒株(稀释度 10 )在牛气管和马肾细胞培养物中引起的细胞病变效应(CPE)从第一到第十代都是稳定的,生物活性为 5.75-6.00 lgTCID 。病毒引起的 CPE 在第 2-3 天出现,在第 5-7 天进一步强烈发展并扩展至细胞单层的 60-80%。疫苗结果可用于农场马匹针对鼻肺炎的免疫接种,马匹应每 2-3 个月免疫两次。

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