兔体中和试验用猪瘟病毒阴、阳性血清国家参考品的研制

Preparation of National Reference Materials for Negative and Positive Antisera of Classical Swine Fever Virus（Neutralization Test in Rabbits）

关键词:  猪瘟病毒  血清  国家参照品  兔体中和试验

Key words:  Classical swine fever virus  antiserum  national reference materials  neutralization test in rabbits

摘要:

为制备和标定猪瘟病毒阴、阳性血清国家参照品（兔体中和试验），本试验制备了猪瘟病毒阴性、弱阳性和强阳性血清，通过过滤、分装、冻干、熔封，获得3亚批候选物，并进行了检验、协作标定和定值。结果显示：物理性状、无菌检验、安全检验、支原体检验、真空度测定、均匀性检验和保存有效期试验均合格；剩余水分均小于4%；特异性检验结果均为口蹄疫、伪狂犬病、猪繁殖与呼吸综合征、猪细小病毒病、牛病毒性腹泻/粘膜病抗体阴性，并且弱阳性和强阳性血清能中和猪瘟兔化弱毒而阴性血清不能中和；残余猪瘟病毒免疫荧光和套式RT-PCR核酸检测结果均为阴性；协作标定定值的兔体中和效价分别为：阴性（200905-1）、1：10_{0.928?0.103（200905-2）、1：102.928?0.103（200905-3）。该候选物可以作为国家参照品。}

Abstract:

To prepare and characterize national reference materials for negative and positive antisera of classical swine fever virus (CSFV) (neutralization test in rabbits), negative, weak and strong positive antisera of CSFV were prepared, collected, filtrated, filled, freeze-dried, and sealed. The three sub-batches of candidates were tested, collaborative studied and characterized. The results were as follows. Character, sterility, safety, mycoplasmas, vacuum degree, homogeneity and storage: the candidates passed all the examinations. Residual moisture: all less than 4%. Specific: antibodies against foot and mouth disease, pseudorabies, porcine reproductive and respiratory syndrome, porcine parvovirus and bovine viral diarrhoea-mucosal disease were all negative; while low weak and strong positive antisera of CSFV could neutralize CSFV (lapinized strain) but negative antisera could not. Remainder of CSFV: IFA and nested RT-PCR results were all negative. Collaborative study and characterization: neutralization titers tested in rabbits were negative (200905-1), 1：10 0.928?0.103 (200905-2) and 1：10 2.928?0.103 (200905-3). It could be concluded that the candidates could be used as national reference materials.