4种结核分枝杆菌皮肤变态反应原的动物试验

Animal test on four kinds of Mycobacterium tuberculosis skin allergen

目的通过不同分枝杆菌致敏豚鼠皮试反应的大小评价4种结核分枝杆菌皮肤变态反应原的诊断效果。方法用结核分枝杆菌和卡介苗分别致敏豚鼠,致敏成功后采用轮圈法于每只豚鼠皮内分别注射TB-PPD、BCGPPD、重组结核分枝杆菌ESAT6-CFP10变态反应原(简称EC)和重组结核分枝杆菌11k Da变态反应原(简称11k Da)4种皮肤变态反应原各0.1 mL。记录注射后24及48 h注射局部红肿反应的纵径和横径,计算其均值。结果在不同分枝杆菌致敏豚鼠中,4种皮试试剂各自24及48 h皮试反应大小差异均无统计学意义(P>0.05)。在结核分枝杆菌致敏豚鼠中,TB-PPD和BCG-PPD皮试反应均为阳性,48 h局部红肿直径分别为(11.1±1.4)和(8.7±2.4)mm,差异有统计学意义(P<0.05);EC和11kDa皮试反应也均为阳性,48 h局部红肿直径分别为(15.4±2.3)和(17.0±1.7)mm,差异无统计学意义(P>0.05),但两者反应强度均高于TB-PPD和BCG-PPD。在卡介苗致敏豚鼠中,TB-PPD和BCG-PPD皮试反应均为阳性,48 h局部红肿直径分别为(11.8±1.3)和(11.4±1.5)mm,差异无统计学意义(P>0.05)。而EC和11kDa皮试反应均为0。结论 EC和11kDa可有效鉴别结核分枝杆菌感染和卡介苗接种,而TB-PPD和BCG-PPD均不能对其进行有效鉴别。

Objective To evaluate the diagnostic effect of 4 kinds of Mycobacterium tuberculosis skin allergen by skin tests in guinea pigs sensitized with various mycobacteria. Methods Guinea pigs were sensitized with M. tuberculosis and BCG respectively,and injected i. d. with TB-PPD,BCG-PPD,recombinant M. tuberculosis ESAT6-CFP10 allergen and recombinant M. tuberculosis 11 k Da allergen,0. 1 ml for each. Longitudinal and transverse diameters of redness and swelling in injection sites were recorded 24 and 48 h after injection,of which the mean was calculated. Results The skin reactions of four allergens 24 and 48 h after injection showed no significant difference in guinea pigs sensitized with various mycobacteria（P〈0. 05）. Both TB-PPD and BCG-PPD elicited positive skin reactions in the guinea pigs sensitized with M. tuberculosis,and the reaction diameters were（11. 1 ± 1. 4） and（8. 7 ± 2. 4） mm 48 h after injection respectively,which showed significant difference（P〈0. 05）. EC and 11 k Da also elicited positive skin reactions,and the reaction diameters were（15. 4 ± 2. 3）and（17. 0 ± 1. 7）mm 48 h after injection respectively,which showed no significant difference（P〈0. 05）. However,the reaction intensities of EC and 11 kDa were higher than those of TB-PPD and BCGPPD. In the guinea pigs sensitized with BCG,both TB-PPD and BCG-PPD elicited positive skin reactions,and the reaction diameters 48 h after injection were（11. 8 ± 1. 3） and（11. 4 ± 1. 5） mm respectively,which showed no significant difference（P〉0. 05）. However,no skin reactions were elicited by EC and 11 kDa. Conclusion Both EC and 11 kDa were effective in identification of M. tuberculosis infection and BCG vaccination,while TB-PPD and BCG-PPD were not.