重组结核分枝杆菌（TB）抗原ESAT-6、CFP-10及ESAT-6/CFP-10的表达、纯化及其在结核病患者检测中的诊断潜力
Expression and Purification of Recombinant Mycobacterium Tuberculosis (TB) Antigens, ESAT-6, CFP-10 and ESAT- 6/CFP-10 and Their Diagnosis Potential for Detection of TB Patients.
作者信息
Hemmati M, Seghatoleslam A, Rasti M, Ebadat S, Mosavari N, Habibagahi M, Taheri M, Sardarian A R, Mostafavi-Pour Z
机构信息
Department of Biochemistry, Recombinant Protein Laboratory, Medical School, Shiraz University of Medical Sciences, Shiraz, Iran.
出版信息
Iran Red Crescent Med J. 2011 Aug;13(8):556-63. Epub 2011 Aug 1.
BACKGROUND
One of the most widely used methods to detect tuberculosis (TB) infection is the tuberculin skin test (TST). The completion of Mycobacterium tuberculosis (M. tuberculosis) genome sequence has led to identification of several antigens that can be utilized for accurate diagnosis and control of TB. The aim of this study was to purify the recombinant M. tuberculosis antigens for the evaluation of their potential in TB diagnosis.
METHODS
The recombinant secretory antigens, ESAT-6, CFP-10 and ESAT-6/CFP-10 were produced by PCR and cloning methods. To investigate antigen specific responses of these recombinant antigens in detection of TB, ex vivo enzyme linked immunospot (ELISPOT) test in 30 clinically diagnosed TB patients was evaluated.
RESULTS
The selected M. tuberculosis antigens were cloned, expressed and purified in Escherichia coli (BL21). ELISPOT assay for detection of TB showed the sensitivity of 93, 90 and 100% for recombinant ESAT-6, CFP-10 and ESAT-6/CFP-10 proteins respectively, which is significantly higher than conventional TST.
CONCLUSION
The recombinant antigens of ESAT-6, CFP-10 and ESAT-6/CFP-10 can be used as an accurate means of detecting TB in Iran.
背景
检测结核感染最广泛使用的方法之一是结核菌素皮肤试验（TST）。结核分枝杆菌（M. tuberculosis）基因组序列的完成导致了几种可用于结核病准确诊断和控制的抗原的鉴定。本研究的目的是纯化重组结核分枝杆菌抗原，以评估其在结核病诊断中的潜力。
方法
通过PCR和克隆方法制备重组分泌抗原ESAT-6、CFP-10和ESAT-6/CFP-10。为了研究这些重组抗原在结核病检测中的抗原特异性反应，对30例临床诊断的结核病患者进行了体外酶联免疫斑点（ELISPOT）试验评估。
结果
所选结核分枝杆菌抗原在大肠杆菌（BL21）中进行了克隆、表达和纯化。用于结核病检测的ELISPOT分析显示，重组ESAT-6、CFP-10和ESAT-6/CFP-10蛋白的敏感性分别为93%、90%和100%，显著高于传统TST。
结论
ESAT-6、CFP-10和ESAT-6/CFP-10的重组抗原可作为伊朗结核病的准确检测手段。
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