中国农业大学学报2024，Vol.29Issue(5)：74-80,7.DOI:10.11841/j.issn.1007-4333.2024.05.07
非洲马瘟病毒VP7蛋白ELISA阳性血清标准样品的初步制备
Preliminary preparation of African horse sickness virus VP7 protein ELISA positive serum standard
摘要
Abstract
The study aimed to prepare a positive serum standard of VP7 protein for the ELISA of the African horse sickness virus,a horse without antibodies against common equine diseases was selected as the target animal,and VP7 protein expressed by the prokaryotic expression system of Escherichia coli was used as the immunogen.The immunogen was mixed with MONTANIDE ISA35 adjuvant in equal volume and emulsified,and then the horse was immunized once every 2 weeks for a total of 3 immunizations,and serum was collected to detect the serum antibody titer by ELISA.When the serum potency was highest,a large amount of blood was collected to isolate the serum and verify its sterility,and the high valence serum was lyophilized.The specificity,sterility,uniformity and stability of the serum were tested according to the methods in the Veterinary Pharmacopoeia of the People's Republic of China.The results showed that:1)Antibodies against African horse sickness could be detected in the immunized horse in the third week after immunization.2)The specificity results showed that antibodies against equine influenza virus,equine herpesvirus type Ⅰ,equine herpesvirus type Ⅱ,equine herpesvirus type Ⅳ,equine arteritis virus,equine infectious anemia virus,equine streptococcus,equine Salmonella abortus,Burkholderia pseudomallei,Babesia caballi and Theileria equi were all negative.The prepared standard serum had a clean background and no common equine pathogenic antibodies.3)There was no growth of stray organisms on the case peptone agar medium,tryptic soy peptone liquid medium and thioglycolate liquid medium,indicating that the prepared serum was free from microbial contamination such as bacteria and fungi.4)The stability test results showed that the prepared serum can be stably stored for 2 months at 4,25 and 37 ℃ and for 6 months at-20 and-80 ℃.In conclusion,this study successfully prepared positive serum standard of VP7 protein for the ELISA of the African horse sickness virus,which laid a theoretical foundation for related research on African horse sickness.关键词
非洲马瘟/阳性血清/ELISA/标准样品Key words
African horse sickness/positive serum/ELISA/standard samples分类
农业科技引用本文复制引用
杨燕,胡哲,郭奎,张泽楠,薛红,黄雪英,沈丹,王晓钧..非洲马瘟病毒VP7蛋白ELISA阳性血清标准样品的初步制备[J].中国农业大学学报,2024,29(5):74-80,7.基金项目
十四五国家重点研发计划(2021YFD1800500) （2021YFD1800500）
