结核潜伏感染诊断新型候选标志物的筛选及临床验证

目的 筛选及临床验证结核潜伏感染(LTBI)新型候选标志物,为研究LTBI诊断新方法提供科学基础。方法 选择2013年1月至2015年12月就诊于遵义医学院附属医院的活动性肺结核患者(ATB组)、LTBI者(LTBI组)及健康人群(H组)血清。小样本分别独立地与包含4262个结核分枝杆菌(MTB)重组蛋白的Mtbprot^TM芯片反应进行初筛,选择差异较为明显的蛋白质,定制成蛋白小芯片,进行较大样本验证,对有效数据进行归一化处理,根据单个蛋白的P值进行评价,进一步通过SPSS统计软件,优化出最优组合。结果 用Mtbprot^TM芯片,经32份血清(H组7份、LTBI组9份及ATB组16份)试验验证,初筛出前100个可能与LTBI相关蛋白质,用这些蛋白质制成小芯片,分别与204份血清(H组44份,LTBI组60份,ATB组100份)进行进一步验证试验,优选出8个蛋白标志物,分别为Rv1860、Rv2031c、Rv1441c、Rv0494、Rv3301c、Rv0351、Rv1821和Rv0280,其组合诊断LTBI的特异性及敏感性分别为90.9%和83.1%。结论 成功筛选鉴定出8个LTBI新型候选蛋白标志物,其组合应用在LTBI诊断和鉴别诊断上具有较大的潜在应用前景。

Screening and clinical verification of novel candidate biomarkers for the diagnosis of latent tuberculosis infection

We aimed to screen and verify specific immunogenic proteins of Mycobacterium tuberculosis (MTB) as the candidate biomarkers, to provide a scientific basis of studying new methods for diagnosing latent tuberculosis infection (LTBI). The serum samples of the participants including the patients with active pulmonary tuberculosis (APTB),LTBI individuals (LTBI)and healthy people (HP) were collected in the Affiliated Hospital of Zunyi Medical College from January 2013 to December 2015. We selected 32 serum samples including 7 HP, 9 LTBI and 16 APTB to react respectively with Mtbprot^TM chip which contained 4 262 recombinant proteins of MTB. Based on the screening results, the first 100 significant difference responsive MTB proteins possibly associated with LTBI were screened out. The 100 proteins were customized into a small Mtbprot^TM protein chip. Then, this chip was respectively hybridized with 204 serum samples including 44 H, 60 LTBI and 100 APTB. The effective data were normalized and evaluated on the P value of their single protein, then, the optimal combination was optimized by SPSS software. We found eight new candidate protein biomarkers associated with LTBI including Rv1860, Rv2031c, Rv1441c, Rv0494, Rv3301c, Rv0351, Rv1821 and Rv0280.The specificity and sensitivity of the diagnosis for LTBI with the combination of the eight proteins were 90.9% and 83.1%, respectively. In conclusion, we have found eight novel candidate protein biomarkers associated with LTBI, of which the composite application has great potential application in diagnosis.