犬细小病毒血凝抑制抗体效价胶体金免疫层析检测方法的建立及应用

试验旨在利用胶体金免疫层析技术建立快速检测犬血清中犬细小病毒（canine parvo virus, CPV）血凝抑制（haemagglutination inhibition, HI）抗体效价的方法，用于CPV疫苗免疫效果评价。采用双抗体夹心法，以抗CPV血凝相关抗原的单克隆抗体制备CPV抗原检测试纸条；将犬血清进行不同比例系列稀释后，分别与定量CPV抗原充分反应，滴入CPV胶体金试纸条，根据试纸条检测线（test line，T线）消失时的血清最高稀释倍数判断血清中CPV抗体的HI效价；用此方法检测86份犬血清样品，并与传统血凝抑制试验方法进行分析比较。结果显示，成功制备CPV抗原检测试纸条，确定了试纸条检测犬血清CPV-HI效价的反应条件和结果判定标准。结果表明，在检测不同稀释倍数犬血清反应后的CPV抗原时，能使试纸条T线消失时的血清最高稀释倍数与HI效价具有正相关性，犬血清最高稀释倍数乘以4即为HI效价；两种方法的符合率达90.7%。本试验初步建立了胶体金试纸条检测CPV血凝抑制效价的方法，为检测CPV-HI效价提供了一种操作简单、快速的试验方法，可用于CPV疫苗免疫效果评价。

Establishment and Application of Colloidal Gold Immune Chromatography Test Method for Detection of Canine Parvovirus Hemagglutination Inhibition Antibody Titer

The study was aimed to use colloidal gold immune chromatography technology to establish a rapid method for detection of canine serum canine parvovirus (CPV) hemagglutination inhibition (HI) titer and CPV vaccine immunization effect assessment.Double antibody sandwich method and monoclonal antibodies of anti-CPV hemagglutination antigen were used to prepare CPV antigen test strip.Canine serum with different proportion respectively was mixed with quantitative CPV antigen for full reaction,then dropped the mixture into the CPV colloidal gold test strip,so according to the highest serum dilution ratios when the test strip line T (line T) vanishes,it was to judge CPV antibodies in serum of the HI titer.This method had been used to detect 86 canine serum samples,at the same time,analyzing and comparing it with traditional hemagglutination inhibition test method.The results showed that the CPV antigen detection test strip was successfully prepared,and the reaction conditions and results of the test strip for detecting the titer of CPV-HI in canine serum were determined.The results indicated that when detecting CPV antigen after the dilution of different ratios of canine serum,the highest serum dilution ratios when the strip line T vanished and the HI titer had positive correlation.The highest dilution ratios of canine serum multiplied by 4 was the HI titer.The results of two methods had 90.7% consistency.This experiment established the colloidal gold immune chromatography test strip for the detection of CPV-HI titers method initially.This CPV-HI detection provided a simple and fast test method for the effect evaluation of CPV vaccine immune.