中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2011年
3期
197-200
,共4页
陆晔%梅建%胡衡%罗继%武洁%沈鑫%桂晓红
陸曄%梅建%鬍衡%囉繼%武潔%瀋鑫%桂曉紅
륙엽%매건%호형%라계%무길%침흠%계효홍
分枝杆菌,结核%蛋白质组学%色谱法,液相%酶联免疫吸附测定
分枝桿菌,結覈%蛋白質組學%色譜法,液相%酶聯免疫吸附測定
분지간균,결핵%단백질조학%색보법,액상%매련면역흡부측정
Mycobacterium tuberculosis%Proteomics%Chromatography liquid%Enzyme-linked immunosorbent assay
目的 探讨人MTB标准株H37Rv与牛分枝杆菌标准株卡介苗株的亚细胞蛋白质组学差异.方法 应用密度梯度离心法分离H37Rv和卡介苗株的细胞壁、细胞膜和细胞质蛋白,利用二维液相色谱分离技术进一步获得H37Rv和卡介苗株亚细胞蛋白质组,采用酶联免疫吸附试验分别检测H37Rv和卡介苗各组分与结核病患者和健康人各5例血清的免疫学反应.组间比较采用t检验.结果 共获得26个特异性引发免疫反应的H37Rv菌株蛋白组分.细胞膜组分M3Fr18与结核病患者的血清免疫学反应的吸收光度(A450值)为(721±3)×10-3,明显高于健康人[(356±6)×10-3],也明显高于卡介苗株细胞膜相应组分与健康人的血清免疫学反应强度[(414±7)×10-3],差异均有统计学意义(t值分别为1.852和1.037,均P＜0.01).细胞膜组分MI0Fr21与结核病患者的血清免疫学反应的A450值为(954±6)×10-3,明显高于健康人[(415±6)×10-3],也明显高于卡介苗株细胞膜相应组分与健康人的血清免疫学反应强度[(315±4)×10-3],差异均有统计学意义(t值分别为2.113和2.550,均P＜0.01).结论 二维液相色谱和酶联免疫吸附试验联合应用,有助于检测人MTB感染的相关抗原蛋白.M3Fr18和M10Fr21蛋白质组分可引起结核病患者的特异性免疫反应,提示其可能是人MTB感染的特异抗原.
目的 探討人MTB標準株H37Rv與牛分枝桿菌標準株卡介苗株的亞細胞蛋白質組學差異.方法 應用密度梯度離心法分離H37Rv和卡介苗株的細胞壁、細胞膜和細胞質蛋白,利用二維液相色譜分離技術進一步穫得H37Rv和卡介苗株亞細胞蛋白質組,採用酶聯免疫吸附試驗分彆檢測H37Rv和卡介苗各組分與結覈病患者和健康人各5例血清的免疫學反應.組間比較採用t檢驗.結果 共穫得26箇特異性引髮免疫反應的H37Rv菌株蛋白組分.細胞膜組分M3Fr18與結覈病患者的血清免疫學反應的吸收光度(A450值)為(721±3)×10-3,明顯高于健康人[(356±6)×10-3],也明顯高于卡介苗株細胞膜相應組分與健康人的血清免疫學反應彊度[(414±7)×10-3],差異均有統計學意義(t值分彆為1.852和1.037,均P＜0.01).細胞膜組分MI0Fr21與結覈病患者的血清免疫學反應的A450值為(954±6)×10-3,明顯高于健康人[(415±6)×10-3],也明顯高于卡介苗株細胞膜相應組分與健康人的血清免疫學反應彊度[(315±4)×10-3],差異均有統計學意義(t值分彆為2.113和2.550,均P＜0.01).結論 二維液相色譜和酶聯免疫吸附試驗聯閤應用,有助于檢測人MTB感染的相關抗原蛋白.M3Fr18和M10Fr21蛋白質組分可引起結覈病患者的特異性免疫反應,提示其可能是人MTB感染的特異抗原.
목적 탐토인MTB표준주H37Rv여우분지간균표준주잡개묘주적아세포단백질조학차이.방법 응용밀도제도리심법분리H37Rv화잡개묘주적세포벽、세포막화세포질단백,이용이유액상색보분리기술진일보획득H37Rv화잡개묘주아세포단백질조,채용매련면역흡부시험분별검측H37Rv화잡개묘각조분여결핵병환자화건강인각5례혈청적면역학반응.조간비교채용t검험.결과 공획득26개특이성인발면역반응적H37Rv균주단백조분.세포막조분M3Fr18여결핵병환자적혈청면역학반응적흡수광도(A450치)위(721±3)×10-3,명현고우건강인[(356±6)×10-3],야명현고우잡개묘주세포막상응조분여건강인적혈청면역학반응강도[(414±7)×10-3],차이균유통계학의의(t치분별위1.852화1.037,균P＜0.01).세포막조분MI0Fr21여결핵병환자적혈청면역학반응적A450치위(954±6)×10-3,명현고우건강인[(415±6)×10-3],야명현고우잡개묘주세포막상응조분여건강인적혈청면역학반응강도[(315±4)×10-3],차이균유통계학의의(t치분별위2.113화2.550,균P＜0.01).결론 이유액상색보화매련면역흡부시험연합응용,유조우검측인MTB감염적상관항원단백.M3Fr18화M10Fr21단백질조분가인기결핵병환자적특이성면역반응,제시기가능시인MTB감염적특이항원.
Objective To compare the proteome of Mycobacterium tuberculosis (MTB) H37 Rv strain with Bovine mycobacterium (Bacillus Calmette-Guerin, BCG) strain at the sub-cellular level. Methods Proteins of the cell wall, membrane and cytolymph of H37 Rv and BCG were extracted by density gradient centrifugation. Sub-cellular proteome of H37 Rv and BCG were analyzed using 2-dimensional liquid chromatography. The immunity reactions of H37 Rv fractions with sera from patients ( n = 5 ) and healthy controls ( n = 5 ), as well as BCG fractions with sera from healthy controls ( n = 5 ) were analyzed by ELISA.Data was analyzed by t test. Results Twenty-six fractions of H37 Rv were found to elicit specific antibody response. Fraction M3Fr18 of H37 Rv reacted with sera from patients. The A450 [ (721 ± 3 ) × 10-3] was higher than that with sera from healthy controls [ ( 356 ± 6) × 10 - 3 ] , as well as the A450 of the corresponding fractions of BCG with sera from healthy controls [ (414 ± 7) × 10-3 ]. The differences between the patient group and the 2 healthy control groups were significant ( t = 1. 852 and 1. 037, all P ＜ 0. 01 ). Moreover,fraction M10Fr21 of H37Rv reacted with sera from the patients. The A450[ (954 ±6) × 10 -3 ] was higher than that with sera from the healthy controls [ (415 ± 6 ) × 10-3 ], as well as the A450 of the corresponding fractions of BCG with sera from the healthy controls [ ( 315 ± 4 ) × 10-3 ]. The differences between the patient group and the 2 healthy control groups were significant ( t = 2. 113 and 2. 550, all P ＜ 0. 01 ).Conclusions The combination of 2-dimensional liquid chromatography and immunology technology is useful in finding antigens associated with MTB infection. Fractions M3Fr18 and M10Fr21 can elicit specific immune reaction among MTB patients, suggesting that they may be specific antigens of MTB infection.
