胶乳凝集试验在家蚕浓核病毒检测中的应用

APPLICATION OF LATEX AGGLUTINATION TEST TO THE DETECTION OF DENSONUCLEOSIS VIRUS OF SILKWORM ( BOMBY X MORI)

应用抗体致敏的胶乳进行凝集试验检测提纯的家蚕浓核病毒,可检测出的最小量为1×10^(-3)OD,比对流免疫电泳法和双向免疫扩散法的灵敏度高100倍,但比斑点免疫结合测定法低10倍;检测蚕体样品时,比对流免疫电泳法检出病毒时间早,检出阳性率高。致敏胶乳的最适IgG量为0.5～1mg/ml,致敏胶乳的最适密度为1％。为排除非特异性反应,致敏胶乳应含有0.1％的BSA;以含0.05％Tween-20的Tris-HCl缓冲液制备检样组织匀浆,静置10～15分钟取上清待检。样品与致敏胶乳混合后10分钟内即可判断结果。

By agglutination tests using antibody-sensitized latex particles for the detection of purified densonucleosis virus (DNV) of silkworm,1×10-8O.D of DNV can be detected. The sensitivity was 100 times higher than that of double immuno-diffusion or counter-immunoelectrophoresis and 10 times lower than that of dot immunobinding assay. Compared with counter-immunoelectrophoresis, this method can detect DNV out earlier in diseased silkworm with higher rate of positive reactions. The most suitable concentration of IgG for sensitizing latex particles is 0.5-1 mg/ml and the most suitable density of sensitized latex particals is 1%. The agglutination reaction can be examined within 10 minutes after mixing the supernatant and sensitized latex. In order to obviate the non-specific agglutination during detection of DNV in silkworm, the concentration of bovine serum albumin should be 0.1% in the sensitized latex suspension. The tissues should be homogenated in Tris-HCl buffer containing 0.05% Tween-20, put aside for 10-15 minutes and then the supernatant used for examination.