Molecular and immunological characterization of Mycobacterium tuberculosis CFP-10, an immunodiagnostic antigen missing in Mycobacterium bovis BCG
Molecular and immunological characterization of Mycobacterium tuberculosis CFP-10, an immunodiagnostic antigen missing in Mycobacterium bovis BCG
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DOI：
10.1128/jcm.38.9.3285-3290.2000
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发表时间：
2000-09-01
影响因子：
9.4
通讯作者：
Houghton, R
中科院分区：
文献类型：
--
作者：
Dillon, DC;Alderson, MR;Houghton, R
为了鉴定可用于活动性结核病（TB）血清学诊断的抗原，我们用来自诊断为活动性肺TB的患者的血清池筛选结核分枝杆菌基因组表达文库。所用血清与重组形式的M缺乏反应性。利用这一策略，我们鉴定了一个基因，以前命名为lhp，它编码一种100个氨基酸的蛋白质，称为培养滤液蛋白10（CFP-10）。免疫印迹分析表明，CFP-10存在于M.结核CFP，表明它可能是分泌或脱落的抗原。纯化的重组CFP-10（rCFP-10）被证明能够在一定百分比的缺乏与r38 kDa的反应性的TB患者中检测特异性抗体，最显著的是在涂片阴性病例中，其中灵敏度从单独的r38 kDa的21%增加到包含rCFP-10的40%。在涂片阳性患者血清中，灵敏度从单独r38 kDa的49%增加到包含rCFP-10的58%。此外，rCFP-10被证明是一种有效的T细胞抗原，在70%的无明显疾病的纯化蛋白衍生物阳性个体中引发外周血单核细胞的增殖反应和γ干扰素产生。对该抗原的应答证明了将rCFP-10纳入用于检测活动性TB感染的多价血清学诊断试验中。rCFP-10还有助于开发能够检测M暴露的重组T细胞诊断试验。结核
In order to identify antigens that may be used in the serodiagnosis of active tuberculosis (TB), we screened a Mycobacterium tuberculosis genomic expression library with a pool of sera from patients diagnosed with active pulmonary TB. The sera used lacked reactivity with a recombinant form of the M. tuberculosis 38-kDa antigen (r38kDa), and the goal was to identify antigens that might complement r38kDa in a serodiagnostic assay, Utilizing this strategy, we identified a gene, previously designated lhp, which encodes a 100-amino-acid protein referred to as culture filtrate protein 10 (CFP-10), The lhp gene is located directly upstream of esat-6, within a region missing in M. bovis BCG, Immunoblot analysis demonstrated that CFP-10 is present in M. tuberculosis CFP, indicating that it is likely a secreted or shed antigen. Purified recombinant CFP-10 (rCFP-10) was shown to be capable of detecting specific antibody in a percentage of TB patients that lack reactivity with r38kDa, most notably in smear-negative cases, where sensitivity was increased from 21% for r38kDa alone to 40% with the inclusion of rCFP-10. In smear-positive patient sera, sensitivity was increased from 49% for r38kDa alone to 58% with the inclusion of rCFP-10, In addition, rCFP-10 was shown to be a potent T-cell antigen, eliciting proliferative responses and gamma interferon production from peripheral blood mononuclear cells in 70% of purified protein derivative-positive individuals without evident disease. The responses to this antigen argue for the inclusion of rCFP-10 in a polyvalent serodiagnostic test for detection of active TB infection. rCFP-10 could also contribute to the development of a recombinant T-cell diagnostic test capable of detecting exposure to M. tuberculosis.