福建农业学报2016，Vol.31Issue(9)：903-907,5.DOI:10.19303/j.issn.1008-0384.2016.09.001
新型鸭呼肠孤病毒σB和σC蛋白间接ELISA方法的建立
Development of Indirect ELISA for Detecting Novel Duck Reovirus UsingσB andσC Proteins as Coating Antigens
摘要
Abstract
An indirect ELISA was developed using the purified recombinant novel duck reovirus (NDRV)σB andσC proteins as coating antigens. The optimized conditions for the methodology were determined to include:concentrations ofσB at 12.5μg·mL-1 andσC at 6.25μg·mL-1 ,a blocking buffer of 15% FBS,serum samples being diluted 80 times,goat anti-duck HRP-IgG being diluted 400 times,and substrate incubation at 37℃ for 5 min.The assay was found to be specific without any cross reaction with antibodies of MDRV,DHV,MPV,or MD-GPV.The coincidence rate between the indirect ELISA coated with the recombinantσB andσC proteins and that coated with NDRVwas 92.5%.It appeared that the newly developed ELISA exhibited satisfactory sensitivity and specificity on measurements,and could be an alternative means for detecting anti-NDRV antibodies.关键词
新型鸭呼肠孤病毒/原核表达/σB蛋白/σC蛋白/间接ELISAKey words
novel duck reovirus/prokaryotic expression/σB protein/σC protein/indirect ELISA分类
农业科技引用本文复制引用
王锦祥,程晓霞,陈少莺,陈仕龙,林锋强,王劭,朱小丽,肖世峰..新型鸭呼肠孤病毒σB和σC蛋白间接ELISA方法的建立[J].福建农业学报,2016,31(9):903-907,5.基金项目
福建省自然科学基金项目（2014J05036) （）
福建省科技计划项目---省属公益类科研院所基本科研专项（2016R1022-11) （）
