中国兽医科学2017，Vol.47Issue(11)：1349-1356,8.DOI:10.16656/j.issn.1673-4696.2017.11.002
K亚群禽白血病病毒抗体间接ELISA检测方法的建立
Indirect ELISA for detection of antibodies against subgroup K avian leukosis virus
摘要
Abstract
To develop an indirect ELISA method for the detection of antibodies against subgroup K avian leukemia virus,the gp85 gene of ALV-K GDFX0603 strain was amplified by PCR,then the prokaryotic expression plasmid pET-GDFX0603-gp85 was constructed.The induction conditions of prokaryotic expression were screened,then the recombinant protein was purified by Ni colum.Using purified recombinant protein as coating antigen,an indirect ELISA method for the detection of antibodies against ALV-K was determined.The results showed that the gp85 recombinant protein was expressed as inclusion bodies,and its size was 53 ku.Western-blot analysis showed that recombinant protein had good immunogenicity.The intra-and inter-assay demonstrated that the coefficient of maximum variation was 10.41％ and 12.57％,respectively.The specific tests showed that there were no cross reactions with the sera against ALV-B,ALV-J,AIV,NDV,IBDV,MDV,REV,Escherichia coli and Salmonella,while it had weak cross reactions with the serum against ALV-A.The results showed that the indirect ELISA method for the detection of antibodies against ALV-K had good reproducibility and better specificity.关键词
K亚群禽白血病病毒/gp85蛋白/原核表达/间接ELISAKey words
ALV-K/gp85 protein/prokaryotic expression/indirect ELISA分类
农业科技引用本文复制引用
袁丽霞,曹伟胜,饶明章,张杰,赵子君,陈建,陈杨一骏,严立福,郑晓翠,谢暮晴..K亚群禽白血病病毒抗体间接ELISA检测方法的建立[J].中国兽医科学,2017,47(11):1349-1356,8.基金项目
国家自然科学基金项目(31672552) （31672552）
广东省科技计划项目(2015A020209145) （2015A020209145）
广东省自然科学基金项目(2016A030313403) （2016A030313403）
广东省家禽产业技术体系疾病控制岗位专家项目(2016LM1114) （2016LM1114）
