猪瘟兔化弱毒间接免疫荧光鉴定方法的建立  

Establishment of an Indirect Immunofluorescence Assay for HCLV Titer Detection

作　　者：牛成明 姚静 杨灵芝 杨琴 马景霞 朱杰 

机构地区：[1]山东滨州沃华生物工程有限公司,山东滨州256606 [2]山东省动物病原微生物工程实验室,山东滨州256606 [3]滨州出入境检验检疫局,山东滨州256603 [4]山东滨州市畜牧兽医局,山东滨州256600

出　　处：《中国动物检疫》2016年第10期86-89,共4页China Animal Health Inspection

摘　　要：为快速有效测定猪瘟疫苗病毒含量,建立了猪瘟兔化弱毒(HCLV)间接免疫荧光试验(IFA)方法,并与兔体定型热试验方法进行了初步比较。试验结果表明,以冷甲醇固定ST细胞,将所选猪瘟活疫苗检验用阳性血清1:40倍稀释、荧光二抗1:32倍稀释时,IFA检测HCLV感染的ST细胞清晰、特异,检测猪伪狂犬病毒、猪流行性腹泻病毒、猪传染性胃肠炎病毒感染细胞阴性。对不同猪瘟疫苗半成品的检测结果显示,IFA测定半数组织感染量(TCID50)与兔体定型热试验测定兔体感染量(RID)具有较好的相关性,其拟合曲线为RID/m L=2.783TCID50/m L+110107.213。An indirect immunofluorescence assay （IFA） was developed for detecting HCLV titer rapidly and effectively by using the positive serum of classical swine fever （CSF） as the first antibody and the fluorescein isothiocyanated （FITC） labeled rabbit anti-pig IgG as the second antibody. Results showed the optimum working concentration of the first antibody was 1：40, and the dilution ratio of second antibody was 1：32. The IFA displayed clear and specific fluorescence in ST cells inoculated with hog cholera lapinized virus （HCLV） utilizing the above mentioned method, while negative results were obtained in ST cells inoculated with pseudorabies virus （PRV）, porcine epidemic diarrhea virus （PEDV） and transmissible gastroenteritis virus （TGEV） . Detection for different CSF semi-finished vaccines showed good correlation between Rabbit infection dose （RID） tested with rabbits and 50% tissue culture infective dose （TCIDso） tested with IFA, and the fitted curve was RID/mL=2.783TCID50/ mL＋110107.213.

关 键 词：猪瘟兔化弱毒(HCLV) 间接免疫荧光试验 兔体感染量 

分 类 号：S851.3[农业科学—预防兽医学]