在感染猪萨佩罗病毒的BHK - 21细胞系中进行免疫细胞化学分析。
Immunocytochemistry assay in BHK-21 cell line infected with Porcine Sapelovirus.
作者信息
Kumari Swati, Singh Rahul, Desingu P A, Ray P K, Taru Sharma G, Saikumar G
机构信息
Division of Pathology, ICAR- Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, 243122, India.
Physiology and Climatology Division, ICAR- Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, 243122, India.
出版信息
Cytotechnology. 2019 Jun;71(3):751-755. doi: 10.1007/s10616-019-00315-4. Epub 2019 Apr 22.
The present study describes an immunocytochemistry (ICC) assay with self-raised hyperimmune sera and a Baby Hamster Kidney-21 (BHK-21) cell line infected with Porcine Sapelovirus (PSV). Sapelovivus/IVRI/SPF-c-6/2015 strain Indian PSV was isolated from the porcine IBRS-2 cell line and investigated for growth on non-porcine cell lines. After two passages, PSV was successfully grown in BHK-21 and produced the same cytopathic effects as in IBRS-2 such as shrinking of cytoplasm, rounding of cells and detachment of cells from the surface of flask within 24 h. For raising of hyperimmune sera, PSV was grown in IBRS-2 cell line up to the required volume and purified by ultracentrifugation. With self-raised hyperimmune sera in laboratory rats, ICC was performed in BHK-21 cells infected with PSV. Positive signals consisted of large granular aggregates of virus in the cytoplasm near the nucleus, suggesting that PSV can infect cell lines other than those of porcine origin.
本研究描述了一种免疫细胞化学（ICC）检测方法，该方法使用自制的超免疫血清以及感染了猪萨佩洛病毒（PSV）的幼仓鼠肾-21（BHK-21）细胞系。从猪IBRS-2细胞系中分离出萨佩洛病毒/IVRI/SPF-c-6/2015株印度PSV，并研究其在非猪细胞系上的生长情况。经过两代传代后，PSV在BHK-21细胞中成功生长，并产生了与在IBRS-2细胞中相同的细胞病变效应，如细胞质收缩、细胞变圆以及在24小时内细胞从培养瓶表面脱落。为了制备超免疫血清，将PSV在IBRS-2细胞系中培养至所需体积，然后通过超速离心进行纯化。在实验室大鼠中使用自制的超免疫血清，对感染PSV的BHK-21细胞进行了ICC检测。阳性信号表现为细胞核附近细胞质中大量的病毒颗粒聚集体，这表明PSV可以感染除猪源细胞系以外的其他细胞系。
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