涂阴肺结核病人血清蛋白组学研究及SHBG蛋白验证

周颖; 李翠萍; 何晓; 李洪涛; 臧宁; 郑艳燕; 何敏

doi:10.16462/j.cnki.zhjbkz.2016.06.006

涂阴肺结核病人血清蛋白组学研究及SHBG蛋白验证

doi: 10.16462/j.cnki.zhjbkz.2016.06.006

周颖¹,
李翠萍¹,
何晓²,
李洪涛³,
臧宁³,
郑艳燕¹,
何敏^1, ,

1. 广西医科大学公共卫生学院劳动卫生与环境卫生学教研室, 广西南宁 530021;
2. 南宁市疾病预防控制中心公共卫生科, 广西南宁 530023;
3. 广西医科大学医学科学实验中心, 广西南宁 530021

基金项目:

广西科技厅资助项目（桂科攻1140003A-2，桂科基0991011）；南宁市科技局资助项目（201109063C）；广西高校人才小高地建设创新团队资助计划（桂教人[2010]38号）

详细信息

作者简介:
周颖(1990-),男,江西宜春人,在读硕士研究生。主要研究方向:环境基因组。

中图分类号: R521
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出版历程
- 收稿日期: 2016-01-21
- 修回日期: 2016-04-25

Discovering candidate serum biomarker of smear-negative pulmonary tuberculosis by proteome quantitative analysis and verifying SHBG

1. Labor Health and Environmental Hygiene Department, School of Public Health, Guangxi Medical University, Nanning 530021, China;
2. Public Health Division, Nanning Center for Disease Control and Prevention, Nanning 530023, China;
3. Medical Scientific Research Center of Guangxi Medical University, Nanning 530021, China

摘要

摘要: 目的利用蛋白质组学方法发现和验证涂阴肺结核的候选血清标志物。方法采用iTRAQ标记结合MALDI-TOF/MS筛选30例健康对照者和30例涂阴肺结核患者血清的差异表达蛋白，通过生物信息学分析差异蛋白之间的相互作用；对差异蛋白SHBG进一步ELISA验证，应用SPSS 20.0统计软件绘制ROC曲线。结果三次独立重复实验一共筛选和鉴定置信度在95%以上蛋白283种；以正常人血清为对照，涂阴肺结核患者血清中有差异表达蛋白31种（P<0.05），其中17个蛋白表达上调，14个蛋白表达下调；蛋白相互作用发现SHBG等12种蛋白处于相互作用网络关键节点；ELISA验证结果显示涂阴肺结核组血清SHBG蛋白浓度高于正常对照组和肺炎组（P<0.001），与蛋白组学筛选和鉴定的结果一致；以SHBG检测值绘制的ROC曲线下的面积是0.92（P<0.001），其诊断涂阴结核病的灵敏度和特异度分别是87.14%和85.11%。结论 iTRAQ标记结合MALDI-TOF/MS技术筛选和鉴定到的12种关键节点蛋白可能是涂阴结核病潜在标志物，SHBG是值得进一步研究的肺结核候选血清标志物。
- 结核,肺 /
- 蛋白质组学 /
- 同位素标记
Abstract: Objective To discover and verify the serum differential expression proteins of smear-negative pulmonary tuberculosis by using a comparative proteomics approach. Methods Serum protein expression profiles from 30 cases of normal healthy controls, and 30 cases of smear-negative pulmonary tuberculosis patients were detected by iTRAQ labeling in combine with MALDI-TOF/MS,and analyzed by bioinformatics software. A proteins interaction diagram was drew,and SHBG was validated by ELISA. ROC was built by SPSS 20.0 software. Results Screened by iTRAQ labeling in combine with MALDI-TOF/MS,the confidential levels of 283 proteins were over 95%. Within 31 significantly differentially expressed proteins, the expression levels of 17 proteins were significantly increased, and 14 proteins were decreased in smear-negative pulmonary tuberculosis patients serum when compared to the normal healthy controls (P<0.05). After analysis on their functional interactions, 12 proteins such as SHBG appeared at the central position of the functional network. SHBG was found to be elevated in smear-negative pulmonary tuberculosis patients when compared to the normal controls and pneumonia patients as examined by ELISA (P<0.001), which was consistent with the iTRAQ result. Examining serum SHBG by ELISA achieved an area under the ROC of 0.92(P<0.001), the sensitivity was 87.14% and specificity was 85.11% in diagnosing smear-negative pulmonary tuberculosis. Conclusions These 12 proteins in the center of the functional network, screened by iTRAQ in combination with MALDI-TOF-MS might be the potential markers of smear-negative pulmonary tuberculosis. SHBG is suggested to be a possible serum candidate biomarker for pulmonary tuberculosis patients.
- Tuberculosis, pulmonary /
- Proteomics /
- Isotope labeling

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参考文献(14)

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