摘要
目的评估11种结核分枝杆菌(Mycobacterium tuberculosis,MTB)抗原用于MTB致敏后豚鼠皮肤试验反应的效果。方法选取18只无特定病原体(specific pathogen free,SPF)级豚鼠(雌性,体质量250~300g),按接种剂量分为高(5μg)、中(0.5μg)、低(0.1μg)3组,每组6只。采用MTB标准株(H37Rv,ATCC27294)致敏豚鼠,致敏成功后豚鼠背部去毛皮内注射结核菌素纯蛋白衍生物(TB-PPD)、重组结核杆菌融合蛋白(EC)和11种MTB抗原(包括重组抗原Rv3872、MPT64、Rv1985c、Rv0222、Rv3117、Rv3120、Rv2346c、Rv3619c、Rv3425、Rv1738和Rv2626c),各0.1ml。分别于注射24h和48h后观察皮肤试验反应,测量硬结平均直径[(横径+纵径)/2],以硬结平均直径≥5mm为阳性。分析11种MTB抗原皮肤试验反应与TB-PPD和EC的差异。结果抗原注射24h后,剂量为5μg时Rv3120、Rv3619c的皮肤试验反应阳性率分别为9/9和8/9;剂量为0.5μg时EC、MPT64的皮肤试验反应均为阳性。TB-PPD(5IU)及EC(0.5μg剂量)、MPT64(0.5μg剂量)、Rv3120(5μg剂量)和Rv3619c(5μg剂量)皮肤试验反应24h硬结平均直径[中位数(四分位数)]均大于反应48h[硬结平均直径分别为:9.00(7.00,11.00)mm和6.50(5.50,8.25)mm、13.00(12.75,14.25)mm和9.00(8.00,9.75)mm、9.50(8.50,12.50)mm和8.50(5.50,9.50)mm、8.50(6.25,9.25)mm和5.00(0.00,5.50)mm、6.50(5.50,8.25)mm和3.50(1.50,4.75)mm],差异均有统计学意义(Z值分别为-2.494、-2.677、-2.207、-2.673、-2.670;P值分别为0.013、0.007、0.027、0.008、0.008)。皮肤试验反应24h时,MPT64(0.5μg剂量)、Rv3120(5μg剂量)和Rv3619c(5μg剂量)与TB-PPD(5IU)硬结平均直径接近,差异均无统计学意义(H值分别为-0.496、0.819和1.714,P值分别为1.000、1.000和0.865);MPT64(0.5μg剂量)与EC(0.5μg剂量)皮肤试验反应情况接近,差异无统计学意义(H=2.288,P=0.221);Rv3120(5μg剂量)和Rv3619c(5μg剂量)硬结平均直径均小于EC(0.5μg剂量),差异均有统计学意义(H值分别为3.795和4.690,P值分别为0.001和0.000)。结论MPT64(0.5μg剂量)和Rv3120(5μg剂量)的豚鼠皮肤试验效果相对较好,对于结核病诊断具有潜在应用价值。
Objective To evaluate the effect of 11 Mycobacterium tuberculosis(MTB)antigens on the evaluation of skin reactions of MTB-sensitized guinea pigs.Methods The 18 specific pathogen free(SPF)guinea pigs(female,weight:250-300 g)were selected and divided into three groups of high dose(5μg),medium dose(0.5μg)and low dose(0.1μg),with 6 guinea pigs per group.The guinea pigs were sensitized with MTB strain(H37Rv,ATCC27294).After successful sensitization,the guinea pigs were intracutaneously injected with TB-PPD,EC and 11 MTB antigens(including recombinant antigen Rv3872,MPT64,Rv1985c,Rv0222,Rv3117,Rv3120,Rv2346c,Rv3619c,Rv3425,Rv1738 and Rv2626c),0.1 ml for each.The transverse and longitudinal diameters of swelling in injection sites were recorded at 24 hand 48 hafter injection,and the average diameter of induration((transverse diameter+longitudinal diameter)/2)was calculated,the average diameter≥5 mm was defined positive.The differences in skin reactions of 11 MTB antigens compared with TB-PPD and EC were analyzed.Results After 24 hof antigen injection,the positive rates of Rv3120 and Rv3619 cwere 9/9 and 8/9,respectively at the dose of 5μg;the skin reactions of EC and MPT64 were all positive at the dose of 0.5μg.In the skin reactions,the average diameters of induration(M(Q1,Q3))of TB-PPD(at the dose of 5 IU),EC(at the dose of 0.5μg),MPT64(at the dose of 0.5μg),Rv3120(at the dose of 5μg)and Rv3619 c(at the dose of 5μg)24 h after injection were all significantly bigger than those 48 hafter injection(the average diameter of induration(9.00(7.00,11.00)mmvs.6.50(5.50,8.25)mm,13.00(12.75,14.25)mmvs.9.00(8.00,9.75)mm,9.50(8.50,12.50)mmvs.8.50(5.50,9.50)mm,8.50(6.25,9.25)mmvs.5.00(0.00,5.50)mm and 6.50(5.50,8.25)mmvs.3.50(1.50,4.75)mm;Zvalues were-2.494,-2.677,-2.207,-2.673 and-2.670,respectively;Pvalues were 0.013,0.007,0.027,0.008 and 0.008,respectively).At the 24 hskin reaction,the average diameter of induration of MPT64(at the dose of 0.5μg),Rv3120(at the dose of 5μg)and Rv3619 c(at the dose of 5μg)were all similar to that of TB-PPD(at the dose of 5 IU),with no statistical difference(Hvalues were-0.496,0.819 and 1.714,respectively;Pvalues were 1.000,1.000 and 0.865,respectively).The sizes of skin reactions between MPT64(at the dose of 0.5μg)and EC(at the dose of 0.5μg)were similar,with no significant difference(H=2.288,P=0.221).The average diameter of induration of Rv3120(at the dose of 5μg)and Rv3619 c(at the dose of 5μg)were both significantly smaller than that of EC(at the dose of 0.5μg)24 hafter injection(H values were 3.795 and 4.690,respectively;P values were 0.001 and 0.000,respectively).Conclusion The skin reaction of MPT64(at the dose of 0.5μg)and Rv3120(at the dose of 5μg)in guinea pigs were relatively better and showed great potential in the diagnosis of tuberculosis.
作者
段惠娟
褚洪迁
孔成成
戴广明
曹廷明
孙照刚
DUAN Hui-juan;CHU Hong-qian;KONG Cheng-cheng;DAI Guang-ming;CAO Ting-ming;SUN Zhao-gang(Translational Medicine Center,Beijing Chest Hospital,Capital Medical University,Beijing 101149,China)
出处
《中国防痨杂志》
CAS
CSCD
2021年第3期255-260,共6页
Chinese Journal of Antituberculosis
基金
国家自然科学基金(81871691)
北京市自然科学基金委-北京市教委联合资助重点项目(21JG0034)。
关键词
分枝杆菌
结核
抗原
皮肤试验
评价研究
Mycobacterium tuberculosis
Antigens
Skin tests
Evaluation studies
