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目的利用血清蛋白组学技术筛选和鉴定耐多药肺结核患者血清差异表达蛋白。方法采用i TRAQ标记结合MALDI-TOF/MS筛选健康体检者、肺炎患者和耐多药肺结核患者血清的差异表达蛋白,并进行生物信息学分析;对差异蛋白FCN3进行MRM相对定量验证。结果一共筛选和鉴定到248种蛋白质,其中置信度在95%以上有92种;以正常对照者和肺炎患者血清为对照,在耐多药肺结核患者血清中发现和鉴定出29种差异表达蛋白(P<0.05),其中16种表达显著增高,13种表达显著降低,FCN3在耐多药肺结核患者血清中表达降低(P<0.05);MRM相对定量验证显示,与正常对照组相比,FCN3在肺炎和耐多药肺结核组中积分峰面积分别为0.818 5±0.037 7和0.378 6±0.020 2,FCN3在耐多药肺结核组表达水平明显降低(P<0.005)。结论 MRM验证FCN3的表达水平与血清蛋白组学筛选的结果一致,FCN3可能是耐多药肺结核潜在的血清标志物。
Objective To screen and identify differentially expressed proteins in patients with multidrug - resistant pulmonary tuberculosis by serum proteomics. Methods The differentially expressed proteins were detected by i TRAQ-tag combined with MALDI-TOF / MS in healthy volunteers, pneumonia patients and MDR-TB patients, and bioinformatics analysis was performed. The differential protein FCN3 was quantitatively validated by MRM. Results A total of 248 proteins were screened and identified, of which 92 were more than 95% confidence level. Serum samples from healthy controls and pneumonia patients were used as controls to detect and identify 29 differentially expressed proteins The expression of FCN3 in the serum of patients with multidrug-resistant pulmonary tuberculosis was significantly lower than that in the normal control group (P <0.05) The integrated peak area of pneumonia and MDR-TB group were 0.818 5 ± 0.037 7 and 0.378 6 ± 0.020 2, respectively. The expression of FCN3 in MDR-TB group was significantly lower (P <0.005). Conclusions MRM results show that the expression level of FCN3 is consistent with the result of serum proteomics screening. FCN3 may be a potential serum marker for multidrug-resistant pulmonary tuberculosis.