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Development of haemagglutination assay for titration of porcine circovirus type 2

文献类型： 外文期刊

作者： Cheng, Haiwei 1 ; Yang, Li 5 ; Cai, Zizheng 6 ; Qiao, Xuwen 1 ; Du, Luping 1 ; Hou, Jibo 1 ; Chen, Jin 1 ; Zheng, Qisheng 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Vet Immunol & Engn, Nanjing 210014, Peoples R China

2.Jiangsu Acad Agr Sci, Natl Res Ctr Engn & Technol Vet Biol, Key Lab Vet Biol Engn & Technol, Minist Agr, Nanjing 210014, Peoples R China

3.Jiangsu Coinnovat Ctr Prevent & Control Important, Yangzhou 225009, Jiangsu, Peoples R China

4.Minist Sci & Technol, Jiangsu Key Lab Food Qual & Safety, State Key Lab Cultivat Base, Beijing, Peoples R China

5.Nanjing Agr Univ, Coll Vet Med, Nanjing 210095, Peoples R China

6.Nanjing Agr Univ, Nanjing 210095, Peoples R China

关键词： PCV2; Titration; Hemagglutination; Chicken red blood cells; Bispecific nanobody

期刊名称：ANALYTICAL BIOCHEMISTRY （ 影响因子：3.365； 五年影响因子：3.049 ）

ISSN： 0003-2697

年卷期： 2020 年 598 卷

页码：

收录情况： SCI

摘要： Porcine circovirus type 2 (PCV2) was one of the most economically important viral pathogens in all the swineproducing countries and often resulted in tremendous economic losses for the swine industry. As PCV2 could not cause cytopathogenic effects while propagated in infected cells, many complicated experiments should be performed to titrate its virus titer. In this study we developed a simple and effective hemagglutination assay for titration of virus titer of PCV2. To develop the hemagglutination assay, a recombinant bispecific nanobody (BsNb) against PCV2 and chicken red blood cells (cRBCs) was constructed based on two nanobodies (NbPCV11 and NbRBC48) which were selected from the non-immunized nanobody library, respectively. The hemagglutination assay was used to titrate the virus titer of PCV2 propagated in cell culture by simple naked-eye observation within 30 min, with the detection limit of 10(4.09) tissue culture infective dose 50 (TCID50)/mL, excellent specificity and reproducibility. Therefore, the hemagglutination assay had potential to be a rapid, reliable, cost-effective, user-friendly qualitative and semi-quantitative tool for titration of virus titer of PCV2 during the vaccine manufacturing process.

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