多价荧光抗体监测外源病毒污染的研究  

Detection of Exogenous Viruses Contamination in Non-Avian Cells with a Multivalent Fluorescent Antibodies Preparation

作　　者：邵振华[1] 田海燕[1] 

机构地区：[1]中国兽药监察所

出　　处：《中国兽药杂志》1997年第2期1-6,T001,共7页Chinese Journal of Veterinary Drug

摘　　要：用猪瘟病毒（ＨＣＶ）、猪细小病毒（ＰＰＶ）、牛病毒性腹泻／粘膜病病毒（ＢＶＤ／ＭＤＶ）、伪狂犬病病毒（ＰｒＶ）、兰舌病病毒（ＢｔＶ）、羊口疮病毒（ＯＶ）等六种病毒，多次同时或先后强化免疫猪，制取六价高免血清；用狂犬病病毒（ＲＶ）免疫兔，制取ＲＶ单价高免血清。分别用异硫氰酸荧光素（ＦＩＴＣ）标记制成荧光抗体（ＦＡ）。并按二者最佳染色价配制成七价（多价）ＦＡ。由于各病毒在其宿主细胞中复制部位和与宿主细胞相互关系各不相同，在荧光显微镜下，多价ＦＡ对其相应病毒的细胞培养物，能显示出各不相同的荧光特点，而得以检测、鉴定。用相应的多、单价高免血清作抑制染色试验，结果各病毒荧光可被一一抑制，表明了多价ＦＡ具有高度的特异性。对各病毒进行ＦＡ细胞培养物半数感染量（ＦＡ－ＴＣＩＤ５０）与常规ＴＣＩＤ５０或ＬＤ５０测定比较试验，结果前者均高于后者，表明了这种荧光抗体制剂同时具有高度的敏感性。应用多价ＦＡ，对非禽源细胞苗生产中外源病毒的污染进行了中间监测，选用了与强毒荧光无区别的７种感染的细胞作阳性参照，保证了生产单位在使用中的安全，对Ａ、Ｂ二厂生产猪瘟苗有干扰的细胞，进行抽样监测８０批，结果１３批污染ＢＶＤＶ（１６．２５％），１?A multivalent fluorescent antibodies preparation was produced for the detection of exogenous viruses contamination in non-avian cells. This is a mixture of a hexavalent fluorescent antibodies preparation and a monovalent preparation. The former antiserum was produced by pigs hyperimmunized wiht the hog cholera virus (HCV) , porcine parvovirus (PPV) , bovine viral diarrhea-mucosal disease virus(BVD/MDV),pseudorabies virus(PrV), bluetonge virus (BtV) and orf virus (OV) , the later one by rabbits hyperimmunized wiht rabies virus (RV) . And each of these two was conjugated with fluorescent isothiocyanate (FITC) separately and then mixtured in an adequate ratio. Under the fluorescent microscopic adamination, every given contaminating virus plaque would be stained in a characteristic feature, on the basis of this character, the exogenous contaminator would be identified. Each of these virus specific fliorescene feature would be inhibited by its specific antiserum in the staining inhibition test. The 50% tissue culture infective dose (TCID50) tested by the fluorescent antibody stain method was higher than that by the routine method.80 samples of cells which produced disruptions in producing live hog cholera vaccine obtained from biologics factories were assayed with this preparation.13 and 12 of them were demonstrated to be contaminated or suspiciously contaminated by BVDV(account for 16.25%) respectively, and anather 3 contaminated by OV (3.8%) . On these results basis, the authors suggest that this preparation is of both high specificity and sensitiveity and of high practicability and reliability.

关 键 词：多价荧光抗体 HCV PPV PRV BTV 药物 病毒污染 

分 类 号：S852.65[农业科学—基础兽医学] S859.2[农业科学—兽医学]