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目的研究重组结核分枝杆菌ESAT6-CFP10蛋白对临床常见致病分枝杆菌及常见环境分枝杆菌致敏豚鼠的皮试反应。方法以鸟分枝杆菌、胞内分枝杆菌、蟾蜍分枝杆菌、堪萨斯分枝杆菌、海分枝杆菌、瘰疠分枝杆菌、戈登分枝杆菌、苏加分枝杆菌、龟分枝杆菌脓肿亚种、偶发分枝杆菌、草分枝杆菌、微黄分枝杆菌、非洲分枝杆菌、牛分枝杆菌、结核分枝杆菌与卡介菌等16株菌株的活菌菌液分别经腹股沟皮下攻击豚鼠,每组5只,3～4周后,分别皮内注射TB-PPD、PPD-B和重组结核分枝杆菌ESAT6-CFP10蛋白各0.1ml,于注射后24、48h观察局部硬结的纵径与横径,根据48h的反应结果进行判定。结果PPD-B对16种分枝杆菌活菌致敏豚鼠的皮试硬结直径均≥5mm,呈阳性,阳性比例均为5/5;TB-PPD对结核、牛、非洲分枝杆菌和卡介菌等活菌致敏豚鼠的皮试硬结直径均≥5mm,呈阳性,阳性比例均为5/5;重组结核分枝杆菌ESAT6-CFP10蛋白对结核、牛和非洲分枝杆菌活菌致敏豚鼠的皮试硬结直径均≥5mm,呈阳性,阳性比例均为5/5;而对鸟、胞内、蟾蜍、堪萨斯、海、瘰疠、戈登、苏加、龟脓、偶发、草、微黄和卡介菌等分枝杆菌活菌、结核分枝杆菌死菌致敏豚鼠的皮试反应均呈阴性,阳性比例均为0/5。结论重组结核分枝杆菌ESAT6-CFP10蛋白能提高结核分枝杆菌感染诊断的特异度并能区别结核分枝杆菌是否存活、卡介菌致敏导致的皮试超敏反应。
Objective To study the skin test response of recombinant Mycobacterium tuberculosis ESAT6-CFP10 protein to common clinical pathogenic Mycobacterium and common environment mycobacterium sensitized guinea pigs. Methods Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium toad, Kansasia mycobacterium, Mycobacterium maritima, Mycobacterium bovis, Mycobacterium Gordonii, Mycobacterium subtilis, Mycobacterium cheloneri Abscess subspecies, sporadic Mycobacterium, Mycobacterium phlei, Mycobacterium maggot, Mycobacterium Africa, Mycobacterium bovis, Mycobacterium tuberculosis and BCG strains of 16 strains of viable bacteria were via the groin Guinea pigs were challenged subcutaneously with 5 mice in each group. After 3 to 4 weeks, 0.1 ml each of TB-PPD, PPD-B and recombinant Mycobacterium tuberculosis ESAT6-CFP10 protein were injected intradermally and were observed at 24 and 48 hours after injection Vertical and horizontal diameter, according to the response of 48h to determine. Results PPD-B of 16 mycobacterial live-sensitive guinea pig skin test induration diameter were ≥ 5mm, were positive, the positive ratio was 5/5; TB-PPD on tuberculosis, cattle, Mycobacterium Africa and card media Strains of sensitized guinea pigs, such as bacteria, were ≥ 5 mm in diameter and positive in positive ratio, all of which were 5/5. Recombinant M. tuberculosis ESAT6-CFP10 protein was sensitive to tuberculosis, bovine and Mycobacterium african mycobacterium sensitized guinea pigs Of the skin test induration diameter are ≥ 5mm, were positive, the positive ratio of 5/5; and birds, intracellular, toad, Kansas, the sea, emu, Gordon, Suga, turtle abortion, sporadic, grass, micro Mycobacterium tuberculosis such as yellow and BCG living bacteria, Mycobacterium tuberculosis sensitized guinea pigs skin test reactions were negative, the positive ratio was 0/5. Conclusion The recombinant Mycobacterium tuberculosis ESAT6-CFP10 protein can improve the specificity of diagnosis of Mycobacterium tuberculosis infection and can differentiate the survival of Mycobacterium tuberculosis, skin test-induced hypersensitivity caused by BCG.