Pradeep Narayan, Veerakyathappa Bhanuprakash, Vinayagamurthy Balamurugan, Madhusudhan Hosamani, Gnanavel Venkatesan, and Raj. Detection of Bluetongue Virus Group-specific Antigen Using Monoclonal Antibody Based Sandwich ELISA[J]. Virologica Sinica, 2010, 25(6): 390-400. doi: 10.1007/s12250-010-3160-y.
A monoclonal antibody (MAb) specific for the bluetongue virus (BTV) group specific antigen (VP7) was characterized for its reactivity with purified virus and recombinant BTV VP7 (rVP7) protein and its suitability for use in the sandwich ELISA. The MAb, designated as 5B5 was specific to VP7 and belongs to IgG2a subclass and was selected for the development of the sELISA in this study. The MAb had a titer of 1:25 with BTV and 1:2 with the rVP7 protein. The sELISA is based on capturing of BTV antigen with VP7 specific MAb followed by detection using BTV polyclonal antiserum raised in rabbits. The assay was evaluated with six cell culture adapted serotypes of BTV that have been isolated from India, 1, 2, 15, 17, 18 and 23. The assay could detect BTV antigen as early as day 8 in blood. It was also successfully applied for the detection of BTV group specific antigen in clinical samples of blood, washed RBCs, buffy coat and plasma. A total of 102 field samples from animals, suspected of being infected with BTV, were tested and 29.42% were positive. The blood samples were also amplified in cell culture which improved the sensitivity of the assay. Results confirmed that the sELISA is rapid and specific.
A. A. El-Kenawy, M. S. El-Tholoth. Sequence Analysis of Attachment Gene of Lumpy Skin Disease and Sheep Poxviruses[J]. Virologica Sinica, 2010, 25(6): 409-416. doi: 10.1007/s12250-010-3150-0.
Egypt, protection of cattle against lumpy skin disease (LSD) was carried out using a sheep poxvirus (Kenyan strain) vaccination strategy. In the present study 15 skin nodules from LSD suspected cows and 5 scab samples from sheep pox (SP) suspected sheep were collected. Hyperimmune rabbit sera to Lumpy skin disease virus (LSDV)/Ismailyia88 strain and sheep pox virus (SPV)/ Kenyan vaccinal strain were prepared. The causative agent in the collected samples was identified using immunoflourescence (IF) and immunoperoxidase techniques. Of the 15 skin nodules suspected of LSD, 10 showed a positive reaction and 3 out of 5 skin scabs suspected of sheeppox were found to be positive. An antigenic correlation between field skin isolate of LSDV, tissue culture adapted LSDV/Ismailyia88 strain, field skin isolate of SPV and SPV/Kenyan vaccinal strain was studied using prepared hyperimmune sera. Also, nucleotide sequence of the PCR amplified attachment gene fragments of field skin isolate of LSDV, tissue culture adapted LSDV/Ismailyia88 strain, field skin isolate of SPV and SPV /Kenyan vaccinal strain were compared. The results revealed that the four used viruses were antigenically identical. Sequence analysis indicated that field skin LSDV isolate is more related to tissue culture adapted LSDV/Ismailyia88 strain than to vaccinal SPV/ Kenyan strain and the skin isolate of SPV is more closely related to field skin isolate of LSDV than to SPV/Kenyan vaccinal strain. Thus, further study should be applied on the advantage of a LSD vaccine prepared from LSDV in protection of cattle against LSD compared to the commonly used sheep pox vaccine.
Yu BAI, rong LI, hua WANG, mei ZHOU, ming ZHAO. 朊蛋白多肽PrP106-126 改变了小鼠小胶质细胞BV-2的 PrP mRNA表达[J]. Virologica Sinica, 2010, 25(6): 440-444. doi: 10.1007/s12250-010-3134-z.
朊病是一种传染性和致死性的神经退行性疾病。朊病病原是一种异常的朊蛋白聚集物。中枢神经系统内的小胶质细胞激活是朊病的一种显著特征。本研究应用实时荧光定量PCR方法检测了朊蛋白多肽PrP106-126对小鼠小胶质细胞BV-2 的PrP mRNA表达影响。结果显示PrP106-126作用BV-2细胞18 h 后PrP mRNA表达水平显著提高,比正常水平提高3倍,作用24 h 后比正常水平提高4.5 倍,且BV-2 细胞的增殖活性在18 h 和24 h 后也相应提高。这些结果首次证明了朊蛋白多肽PrP106-126 提高了小胶质细胞的PrP mRNA 表达水平,表明小胶质细胞在朊病毒致病过程中可能具有关键作用。