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一种鸡抗猪丁型冠状病毒阳性血清的制备方法
李晶梅,张飞雁,王焕君,柏娇,于义娟,王碧群,朱薇,郑良益,李婷婷,冯钊,石宝兰,漆世华,谢红玲
0
(国药集团动物保健股份有限公司)
摘要:
为研究能否用鸡制备出猪丁型冠状病毒(PDCoV)阳性血清。将PDCoV用BEI灭活制备抗原,抗原与矿物油佐剂乳化制备疫苗,疫苗多次免疫SPF鸡,采集血清,检测血清中和效价、将血清用于IFA检测PDCoV。血清中和效价随免疫次数增加逐渐增高,4免后14日的中和效价平均值为1300;中和效价1000左右的血清混合作为阳性血清,1000倍稀释可用于IFA检测PDCoV。实验说明,用鸡制备PDCoV阳性血清的方法成立,制备的阳性血清可用于中和实验、IFA检测PDCoV。
关键词:  猪丁型冠状病毒  阳性血清  中和效价  IFA
DOI:
投稿时间:2022-03-18修订日期:2022-08-04
基金项目:
The preparation of positive porcine deltacoronavirus serum by chicken
(Sinopharm Animal Health Corporation Ltd.)
Abstract:
In this study, we immunized SPF chickens with the vaccines of porcine delta coronavirus (PDCoV) in order to investgate the possibility of prepare the sera against PDCoV by chickens. PDCoV was inactivated with BEI, the inactivated viruses and mineral oil adjuvants emulsified to prepare vaccines, The SPF chickens were immunized with the vaccines multiplies times, sera from immuned chickens was collected, the IFA(Indirect immunofluorescence assay) and serum neutralization test( SNT) were used to detect the antibodies of PDCoV, and neutralizing antibody titer of sera gradually increases due to the increase of immunization times,14 days after the fourth immunication the average neutralizing antibody titer was 1:1300; sera was 1:1000 diluted to the detection of PDCoV by IFA. This study shows that the method of preparing PDCoV positive sera with chicken was feasible, and the prepared positive sera can be used for neutralization and IFA of PDCoV.
Key words:  porcine delta coronavirus  positive serum  neutralizing antibody titer  IFA

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