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目的 :结核分枝杆菌重组Ag85A抗原诱发豚鼠迟发型超敏 (DTH)反应。Ag85A剂量、观察时间对DTH的影响。方法 :常规皮肤实验法 :灭活H37Rv全菌体致敏豚鼠 ,皮肤试验以生理盐水为阴性对照 ,5IUPPD标准品为阳性对照 ,0 .1、0 .2、0 .4、0 .6、0 .8及 1.0 μg重组抗原分别于背部皮内注射 ,注射后 2 4、4 8及 72h分别测量红肿或硬结反应横、纵直径 (mm) ,取两者的平均值。皮肤试验轮圈法 :生理盐水、5IUPPD及 0 .4、0 .6、0 .8、1.0 μgAg85A抗原 ,以轮圈法注射于同一只豚鼠背部一侧 ,每批 6只 ,共 3批。于注射后 2 4、4 8、72h分别测量红肿或硬结反应横、纵直径 (mm) ,取两者的平均值。结果 :常规皮肤试验 0 .1、0 .2 μg重组Ag85A抗原诱发DTH的平均直径与PPD相比有显著差异 (P <0 .0 5 ) ,而 0 .4、0 .6、0 .8、1.0重组Ag85A抗原诱发豚鼠DTH的平均直径与PPD相比 ,无显著差异 (P >0 .0 5 ) ,2 4h平均直径显著高于 4 8h平均直径 (P <0 .0 5 ) ,2 4h测量效果较好。轮圈法皮肤试验排除个体差异后结果与常规皮肤试验结果一致。结论 :从我们的实验结果来看重组Ag85A抗原诱发豚鼠DTH反应可以代替PPD
Objective: Mycobacterium tuberculosis recombinant Ag85A antigen induced guinea pig delayed-type hypersensitivity (DTH) reaction. Ag85A dose, the impact of observation time on DTH. Methods: The conventional skin test: inactivated H37Rv whole body sensitized guinea pigs, skin test with saline as a negative control, 5IUPPD standard as a positive control, 0 .1,0 .2,0 .4,0. .8 and 1.0 μg recombinant antigens were injected intradermally on the back, respectively. The mean and transverse diameters of the inflamed or indurated reaction were measured at 2, 4, 8 and 72 hours after injection, respectively. Skin test Routine method: saline, 5IUPPD and 0 .4,0 .6,0 .8,1.0 μgAg85A antigen, the same way to the guinea pig back side injection, each batch of 6, a total of 3 batches. Measured at 24,48,72h after injection, respectively, horizontal swelling and induration reaction, vertical diameter (mm), whichever is the average. Results: The average diameter of DTH induced by 0.1 microg / ml recombinant Ag85A antigen in routine skin test was significantly different from that of PPD (P <0.05) 1.0 The average diameter of DTH induced by recombinant Ag85A antigen in guinea pigs was not significantly different from that in PPD (P> 0.05), the average diameter at 24 hours was significantly higher than the mean diameter at 48 hours (P <0.05) The effect is better. Routine skin test to exclude individual differences in the results with the conventional skin test results. CONCLUSIONS: From our experimental results, recombinant Ag85A antigen-induced guinea pig DTH response can replace PPD