摘要
用硫酸铵盐析法和蔗糖梯度离心法，从犬瘟热病毒人工感染犬的肝、脾中浓缩提纯犬瘟热病毒，以其免疫家兔，制备兔抗犬瘟热病毒免疫血清，再经硫酸铵沉淀与ＱＡＥ－葡聚糖凝胶Ａ５０层析提取ＩｇＧ，于４℃低速搅拌标记异硫氰酸荧光素，制成兔抗犬瘟热病毒抗原荧光抗体。用上述荧光抗体检测２１只犬瘟热病毒人工感染犬和３５只自然感染犬的１１６份白细胞，抗原阳性１０９份，而健康犬、传染性肝炎犬、犬细小病毒肠炎犬的３７份血液白细胞，全部为抗原阴性。另外，用同样染色法检查了２５份犬瘟热死亡犬脾细胞涂片，有２３份为抗原阳性；而健康犬、传染性肝炎犬和犬细小病毒肠炎犬的１７份脾细胞涂片无一为抗原阳性。对１４１份犬瘟热病犬标本，用电镜或病毒分离证明为犬瘟热病毒抗原阳性的有１３８份，直接ＦＡ染色法检查也为抗原阳性的有１３２份，敏感性为９５．７％；经电镜检查或病毒分离证明为抗原阴性的５７份标本，直接ＦＡ染色检查全部为阴性，特异性为１００％。用直接ＦＡ染色法和以细胞病变（ＣＰＥ）为感染指标的ＴＣＩＤ５０检测法对３５份犬瘟热病毒鸡胚成纤维细胞（ＣＥＦ）培养物的毒力进行检测，结果分别平均为４．８５和４．７２ｌｏｇ１０／０．１ｍＬ但直接ＦＡ染色法比ＣＰＥ客观?
The CDV
was purified and concentrated from the livers and spleens ofdogs infected experimentally with
this virus by ammonium sulfate salting-out and su-crose gradient centrifugation, The
hyperimmune serum against CD was prepared by im-munizing the rabbits with CDV and their
IgG was extracted by ammonium sulfate salt-ing-out and QAE-Sephadex A50 chromatography。
The preparation of fluorescent anti-body was done at 4℃。 The leukocytes were collected from
anticoagulative blood of dogsbeing detected and the smears were made and stained with FA for
examination。 Among116 leucocyte samples from 21 dogs experimentally infected with CDV
and 35 naturallyinfected dogs, 109 samples were positive, whereas 37 samples from healthy
dogs anddogs infected with infectious canine hepatitis and canine parvoviral enteritis were aIl
neg-ative。 Besides, 23 out of 25 smears made of the spleens of the dogs succumb to
distem-per were positive; while among 17 smears made of spleens of healthy dogs and dogs
withinfectious canine hepatitis and canine parvoviral enteritis, none was DOsitive。 141
sam-Ples from distemper-infected dogs were examined by electron microscopy and virus
isola-tion, 138 samples were positive, However,the positive cases verified by FA stainingwere
132, the sensitivity rate was 95.7%。 The negative specimens examined by
electronmicroscopy and virus isolation were all negative by FA technique, the specificity
ratewas 100%, 35 strains of CDV multiplicated in chicken embryo fibroblast cultures
weretitrated simultaneously by FA staning and the cytopathic effect method , the
averageTCID50 were 4.85 and 4.72 log 10/0.1 mL respectively。
出处
《中国兽医学报》
CAS
CSCD
1994年第2期146-150,共5页
Chinese Journal of Veterinary Science
关键词
荧光抗体
犬瘟热
诊断
犬病
fluorescent
antibody
canine distemper
diagnosis
