Lele Xu , Zhihao Chen , Haoyang Gong , Xiuxiu Pei , Yiyao Zhu , Yuchen Lu , Yumiao Wang , Shifa Nan , Yupeng Yin , Qin Zhao , Yunpeng Fan , Yani Sun , Shuqi Xiao
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引用次数: 0
摘要
猪圆环病毒 2 型 (PCV2) 在全球养猪场的发病率很高。PCV2 疫苗是预防和控制 PCV2 的重要手段。PCV2 疫苗的质量控制主要基于动物试验和中和抗体滴定等检测技术。测定疫苗中有效蛋白的含量来衡量疫苗的效力是传统方法的一种很好的替代方法,可以大大加快疫苗的研发速度和检测时间。本研究筛选了一种不仅能有效识别外源表达的 PCV2 Cap 蛋白,而且能有效识别 PCV2 病毒的单克隆抗体(mAb)。利用这种能特异性识别 PCV2 Cap 的 mAb 开发了双抗体夹心 ELISA(DAS-ELISA)。该方法检测到的最低蛋白质含量为 3.5 纳克/毫升。该方法可用于 PCV2 灭活疫苗和亚单位疫苗的质量控制,检测结果与小鼠动物实验结果一致。该方法具有操作简单、灵敏度高、特异性强、适用范围广等优点。它可以检测各类 PCV2 疫苗的有效抗原帽蛋白含量,不仅缩短了疫苗检验时间,而且节约了成本。
Development a high-sensitivity sandwich ELISA for determining antigen content of porcine circovirus type 2 vaccines
Porcine circovirus type 2 (PCV2) is intensely prevalent in global pig farms. The PCV2 vaccine is an important means of preventing and controlling PCV2. The quality control of PCV2 vaccines is predominantly based on detection techniques such as animal testing and neutralizing antibody titration. Measuring the content of effective proteins in vaccines to measure vaccine efficacy is an excellent alternative to traditional methods, which can greatly accelerate the development speed and testing time of vaccines. In this study, we screened a monoclonal antibody (mAb) that can effectively recognize not only the exogenous expression of PCV2 Cap protein but also PCV2 virus. The double antibody sandwich ELISA (DAS-ELISA) was developed using this mAb that specifically recognize PCV2 Cap. The minimum protein content detected by this method is 3.5 ng/mL. This method can be used for the quality control of PCV2 inactivated vaccine and subunit vaccine, and the detection results are consistent with the results of mice animal experiments. This method has the advantages of simple operation, good sensitivity, high specificity and wide application. It can detect the effective antigen Cap protein content of various types of PCV2 vaccines, which not only shorten the vaccine inspection time but also save costs.
期刊介绍:
The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery.
The methods may include, but not limited to, the study of:
Viral components and morphology-
Virus isolation, propagation and development of viral vectors-
Viral pathogenesis, oncogenesis, vaccines and antivirals-
Virus replication, host-pathogen interactions and responses-
Virus transmission, prevention, control and treatment-
Viral metagenomics and virome-
Virus ecology, adaption and evolution-
Applied virology such as nanotechnology-
Viral diagnosis with novelty and comprehensive evaluation.
We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.