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目的 :研究结核分枝杆菌重组 16 0 0 0蛋白 (rPA16 )诱发家兔产生抗体的能力并评价其血清学诊断的价值。方法 :家兔按注射的抗原及佐剂的不同分组 ,皮下多点注射分别进行免疫。 6次免疫后采血 ,双向琼脂扩散试验和酶联免疫吸附试验 (ELISA)跟踪兔血清抗体效价的时间效应并检测兔血清与所试各类分支杆菌PPD有无交叉反应 ;同时检测rPA16与各分支杆菌PPD的免疫羊血清的反应 ,并用rPA16抗原检测人血清标本 ,分析该抗原的临床检测的敏感性和特异性。结果 :兔血清抗体效价结果显示 :5个月后 ,加佐剂的抗原组的兔免疫血清仍能检测到抗体 ,不加佐剂组 2个月后血清抗体检测即为阴性。ELISA检测兔免疫血清比双向琼脂扩散方法要更敏感 ,rPA16抗原加佐剂引起的兔血清抗体滴度最高可达 1∶6 4 0 0 ,不加佐剂组的血清抗体滴度仅为 1∶4 0 0。ELISA法分析抗rPA16的兔血清的特异性表明其特异性较好 ,仅与牛、人型结核分支杆菌的PPD呈阳性反应 ,与其他所试分支杆菌PPD反应均为阴性。同时又分析了rPA16抗原的特异性 ,与人型PPD相比 ,该抗原只与所试各类分支杆菌、BCG、牛分支杆菌免疫的血清呈阳性反应 ,而人型PPD则基本上与结核分支杆菌免疫的血清均呈阳性反应。ELISA检测血清标本结果 :肺结核病组中rPA16和PPD检测敏感性?
Objective: To investigate the ability of Mycobacterium tuberculosis recombinant protein 160 (rPA16) to induce antibody production in rabbits and to evaluate its serological diagnosis. Methods: Rabbits were injected with different doses of antigen and adjuvant, and subcutaneous injections were performed separately for immunization. After six times of immunization, the blood samples were collected by two-way agar diffusion assay and enzyme-linked immunosorbent assay (ELISA) to measure the time effect of rabbit serum antibody titers and to detect the presence or absence of cross-reaction between rabbit serum and all tested mycobacteria PPDs. Mycobacterium PPD immunized sheep serum was detected, and rPA16 antigen was used to detect human serum samples. The sensitivity and specificity of the antigen for clinical testing were analyzed. Results: The titer of rabbit serum antibody showed that the antibody could still be detected in the rabbit serum with adjuvant antigen after 5 months. The serum antibody was negative after 2 months without adjuvant. ELISA rabbit serum immune sera than two-dimensional agar diffusion method is more sensitive to rPA16 antigen adjuvant rabbit serum antibody titer up to 1: 640, without adjuvant serum antibody titer of only 1: 40 0. The specificity of rabbit anti-rPA16 rabbit serum by ELISA analysis showed that the specificity was good, which was only positive with PPD of cattle and human Mycobacterium tuberculosis, but negative with other tested mycobacterium PPD. At the same time, the specificity of rPA16 antigen was also analyzed. Compared with human PPD, this antigen showed positive reaction only with all kinds of mycobacteria, BCG and M. bovis immunized sera, while human PPD was basically similar to tuberculosis branch Bacillus immune serum were positive. ELISA detection of serum samples results: pulmonary tuberculosis group rPA16 and PPD detection sensitivity?