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目的探讨早期分泌性抗原靶6(early secretory antigenic target-6,ESAT-6)、结核菌素纯蛋白衍生物(purified protein derivative,PPD)联合进行血清学诊断结核病的效果。方法 ESAT6基因被插入到原核表达载体pGEX-5T中,与谷胱甘肽S-转移酶(glutathione S-transferase,GST)融合进行表达。表达的GST-ESAT6融合蛋白经亲和层析和分子筛进行纯化。针对64例结核患者血清和40例健康人血清,进行了ESAT6、PPD抗体反应性实验,评估二者血清学诊断的敏感性和特异性。结果融合表达的ESAT6蛋白保持天然的免疫原性,在Western blot实验中,可以被结核病人血清所识别。PPD抗体诊断敏感性是84.4%,特异性是65%。ESAT6抗体诊断敏感性是68.75%,特异性是95%。ESAT6、PPD-酶联免疫吸附法(enzyme-linked immuno sorbent assay,ELISA)联合诊断,敏感性是93.4%,特异性是95%。结论 ESAT6、PPD抗体联合判断可以提高结核病诊断的准确性。
Objective To investigate the effect of early secretory antigenic target-6 (ESAT-6) and purified protein derivative (PPD) in serological diagnosis of tuberculosis. Methods The ESAT6 gene was inserted into prokaryotic expression vector pGEX-5T and fused with glutathione S-transferase (GST) for expression. The expressed GST-ESAT6 fusion protein was purified by affinity chromatography and molecular sieves. For 64 cases of tuberculosis patients serum and 40 healthy human serum, ESAT6, PPD antibody reactivity test to assess the sensitivity and specificity of the two serological diagnosis. Results The expressed fusion protein ESAT6 retained its natural immunogenicity and was recognized by TB patient serum in Western blot. PPD antibody diagnostic sensitivity was 84.4%, specificity was 65%. The ESAT6 antibody has a diagnostic sensitivity of 68.75% and a specificity of 95%. ESAT6, PPD-ELISA (enzyme-linked immuno sorbent assay, ELISA), the sensitivity was 93.4%, the specificity was 95%. Conclusion The combination of ESAT6 and PPD antibody can improve the accuracy of diagnosis of tuberculosis.