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11种结核分枝杆菌抗原皮肤试验反应效果评价

Evaluation of the Effect of Skin Reaction on 11 Mycobacterium tuberculosis Antigens
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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 dee,SPF)级豚鼠(雌性,体质量250~300 g),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0.1 ml。分别于注射24 h和48 h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>5 mm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24 h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(5 IU)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24 h硬结平均直径[中位数(四分位数)]均大于反应48 h[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、8.50(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)。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。 【Abstract】Objective To evaluate the effect of 11 Mycobacterium tuberculasis(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 ino tree 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 stain(H37Rv.ATCC27294).After successful sensitization,the guinea pigs were intracutancously injected with TB-PPD.EC and 11 MTB antigens(including recombinant antigen Rv3872,MPT64,Rv1985c,Rv0222,Rv3117,Rv3120,Rv2346c,Rv3619c,Rv3425,Rv1738 and Rv 2626c)0.1 ml for cach.The transverse and longitudinal diameters of swelling in injection sites were recorded at 24 h and 48 h afer injection,and the average diamererof induration [(transverse diameter+Hlongitudinal diamcter)/2]was calculated,the average diameter 2s mm was defined positive.The differences in skinreactions of 11 MTB antigens compared with TB-PPD and EC were analyzed.Results After 24 h of antigen injection.the positive rats of Rv3120and Rv3619e were 9.9 and 89,respectively at the dose of 5μg;the skin reactions of EC and MPT64 were all positive at the dose of 0.5μg.In theskin reactions.the average diameters of induration(M(Q_(1),Q_(3)))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 doseof 5μg)and R3619c(at the dose of 5μg)24 h after injection were all significantly bigger than those 48 h after injection(the average diameter of induration 9.00(7.00.11.00)mm vs.6.50(5.50,8.25)mm,13.00(12.75,14.25)mm vs.9.00(8.00.9.75)mm.9.50(8.50.12.50)mm vs.8.50(5.510,9.510)mm,8.50(6.25,9.25)mm vs.5.00(0.00.5.50)mm and 6.50(5.50.8.25.)mm vs.3.10(1.50.4.75)mm;Z valueswere-2.494,-2.677,-2.207,-2.673 and-2.670,respectively;P values were 0.013,0.007,0.027,0.008 and 0.08,respectively).At the 24 h skin reaction,the average diameter of induration of MPT64(at the dose of 0.5 pe),Rv3120(at the dose of 5μg)and Rv3619c(at the dose of 5μg)were all similar to that of TB-PPD(at the dose of 5IU),with no statistical difference(H values were-0.496,0.819 and 1.714,respectively;P values were 1.0O0,1.00 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 difrence(H=2.288,P=0.221).The average diameter of induration of Rv3120(at the dose of 5μg)and Rv3619e(at the dose of 5μg)were both significantly smalr than that of EC(at the dose of 0.5 pg)24 h after 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 pis 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 Huijuan;Chu Hongqian;Kong Chengcheng;Dai Guangming;Cao Tingming;Sun Zhaogang(Translational Medicine Center,Beijing Chest Hospital,Capital Medical University,Beijing 101149,China)
出处 《结核病与胸部肿瘤》 2021年第2期109-114,共6页 Tuberculosis and Thoracic Tumor
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 Mycobacteriunx ruberculosis Antigens Skin tcsts Evaluation studies
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