血凝试验和反向间接血凝试验定量检测PCV2 Cap抗原的研究

Study on Quantitative Detection of PCV2 Cap Antigen by Hemagglutination Test and Reverse Indirect Hemagglutination Test

关键词:  PCV2 Cap抗原  反向间接血凝  血凝  红细胞

Key words:  PCV2 Cap antigen  reverse indirect hemagglutination  hemagglutination  erythrocyte

摘要:

为研究能否用血凝和反向间接血凝方法定量检测重组杆状病毒表达的PCV2 Cap抗原,分别使用不同稀释液和不同的动物红细胞测定PCV2 Cap抗原的凝集效价,同时使用PCV2 Cap单抗致敏绵羊红细胞,检测PCV2 Cap抗原、空杆状病毒、CSFV等的反向间接血凝效价(RIHA效价)。结果显示,用猪、鸡、绵羊红细胞在不同稀释液条件下测得的同一批次PCV2 Cap抗原凝集效价为3 log2～9 log2,说明PCV2 Cap抗原可凝集猪、鸡、绵羊红细胞,但红细胞不同、稀释液不同,测得的PCV2 Cap抗原凝集效价也不同。使用PCV2 Cap单抗致敏绵羊红细胞,测得的PCV2 Cap抗原RIHA效价为14 log2,且纯化后和未纯化的PCV2 Cap抗原的RIHA效价均与ELISA定量检测结果有平行关系；空杆状病毒、CSFV、PRRSV、PRV、PEDV、TGEV的RIHA效价均小于1 log2。说明反向间接血凝方法可定量检测重组杆状病毒表达的PCV2 Cap抗原,该方法敏感、特异,适用于疫苗生产中的PCV2 Cap抗原的定量检测。

Abstract:

To study whether PCV2 Cap antigen expressed by recombinant baculovirus can be quantitatively tested by hemagglutination (HA) and reverse indirect hemagglutination (RIHA), the HA titers of PCV2 Cap antigen were tested by different animal erythrocytes with different dilutions, and the RIHA titer of PCV2 Cap antigen, empty baculovirus, CSFV, etc were tested by PCV2 Cap monoclonal antibody sensitized sheep erythrocytes. The HA titers of the same batch of PCV2 Cap antigen were 3 log2～9 log2 tested by different dilutions, which showed that PCV2 Cap antigen can agglutinate the erythrocytes of pig, chicken and sheep; the HA titer of PCV2 Cap antigen varies with different erythrocytes and dilutions. The RIHA titer of PCV2 Cap antigen was 14 log2, tested by PCV2 Cap monoclonal antibody sensitized sheep erythrocytes. The RIHA titers of purified and unpurified PCV2 Cap antigen were parallel to the ELISA results. The RIHA titers of empty baculovirus, CSFV, PRRSV, PRV, PEDV and TGEV were less than 1 log2, which showed that RIHA can quantitatively detect PCV2 Cap antigen expressed by recombinant baculovirus, and the method is sensitive, specific, and suitable for the quantitative detection of PCV2 Cap antigen in vaccine production.