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检测猪细小病毒血清抗体乳胶凝集试验方法的建立及初步应用
Establishment of Latex Agglutination Test to Detect Serum Antibody against Porcine Parvovirus(PPV)
【摘要】 将猪细小病毒在IBRS-2细胞上同步培养增殖,待出现细胞病变后,反复冻融,收获病毒液,用甲醛灭活。随后用经硫酸胺沉淀、透析后浓缩的细小病毒液进行方阵滴定,选择致敏乳胶的最佳条件,制成细小病毒乳胶凝集试验(LAT)抗原。抗原与细小病毒阳性血清反应出现肉眼可见的凝集颗粒,而与生理盐水、PBS、犊牛血清、猪瘟、衣原体、口蹄疫、伪狂犬病、弓形体及萎缩性鼻炎等阳性血清不出现凝集现象。用所建立的细小病毒乳胶凝集试验(LAT)与血凝抑制试验检测了203份猪血清,其中血凝抑制抗体阳性为161份,阴性为42份,阳性率为79.31%;乳胶凝集抗体阳性为150份,阴性为53份,阳性率为73.89%,两种方法阳性符合率为93.16%,经统计学检验P>0.05,两者差异不显著。结果表明,乳胶凝集试验可以作为临床上大量血样进行血凝抑制试验前的初筛,具有简便、准确的优点,在检测细小病毒(PPV)抗体上具有较好的应用前景。
【Abstract】 Porcine parvovirus(PPV) was cultivated on IBRS-2 cell, after CPE was observed, the PPV was harvested, frozen and thawed, then inactivated with Formalin, precipitated with (NH4 )2S4., finally dialysed and concentrated. The most suitable conditions for preparing antigen for PPV latex agglutination test (LAT) were determined by titration. After LAT antigen reacted with PPVpositive serum,agglutination Particles could be observed clearly by naked eyes. It was not the case with normal saline solution, phosphate saline buffer, newborn calf serum, the POSitive sera of Hog Cholera, Chlamydia, FMDV, pseudorabies, Toxoplasmasis, Atrophic Rhinitis, Among 203 serum samples tested by HI test and LAT, the positive number was 161 and 151 respectively. The LAT and HI test shared the same results up to 93. 78%. By statistical analysis, no differentiation between the two methods. The result revealed that LAT could be applied in practice to check PPV antibody as the screening method before HI test.
【Key words】 PPV; Antibody detection; Latex agglutination test; Hemagglutination Inhibition test;
- 【文献出处】 中国预防兽医学报 ,CHINESE JOURNAL OF PREVENTIVE VETERINARY MEDICINE , 编辑部邮箱 ,1999年06期
- 【分类号】S852.65
- 【被引频次】73
- 【下载频次】521