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Int J Biol Sci 2016; 12(2):246-256. doi:10.7150/ijbs.13805

Research Paper

A Group of Novel Serum Diagnostic Biomarkers for Multidrug-Resistant Tuberculosis by iTRAQ-2D LC-MS/MS and Solexa Sequencing

Chong Wang1, Chang-Ming Liu1, Li-Liang Wei2, Li-Ying Shi3, Zhi-Fen Pan4, Lian-Gen Mao1, Xiao-Chen Wan3, Ze-Peng Ping1, Ting-Ting Jiang1, Zhong-Liang Chen1, Zhong-Jie Li1, Ji-Cheng Li1✉

1. Institute of Cell Biology, Zhejiang University, Hangzhou 310058, P.R. China
2. Department of Respiratory Medicine, The Sixth Hospital of Shaoxing, Shaoxing 312000, P.R. China
3. Department of Clinical Laboratory, Zhejiang Hospital, Hangzhou 310013, P.R. China
4. Department of Tuberculosis, The First Hospital of Jiaxing, Jiaxing 314001, P.R. China

Abstract

The epidemic of pulmonary tuberculosis (TB), especially multidrug-resistance tuberculosis (MDR-TB) presented a major challenge for TB treatment today. We performed iTRAQ labeling coupled with two-dimensional liquid chromatography-tandem mass spectrometry (2D LC-MS/MS) and Solexa sequencing among MDR-TB patients, drug-sensitive tuberculosis (DS-TB) patients, and healthy controls. A total of 50 differentially expressed proteins and 43 differentially expressed miRNAs (fold change >1.50 or <0.60, P<0.05) were identified in the MDR-TB patients compared to both DS-TB patients and healthy controls. We found that 22.00% of differentially expressed proteins and 32.56% of differentially expressed miRNAs were related, and could construct a network mainly in complement and coagulation cascades. Significant differences in CD44 antigen (CD44), coagulation factor XI (F11), kininogen-1 (KNG1), miR-4433b-5p, miR-424-5p, and miR-199b-5p were found among MDR-TB patients, DS-TB patients and healthy controls (P<0.05) by enzyme-linked immunosorbent assay (ELISA) and SYBR green qRT-PCR validation. A strong negative correlation, consistent with the target gene prediction, was found between miR-199b-5p and KNG1 (r=-0.232, P=0.017). Moreover, we established the MDR-TB diagnostic model based on five biomarkers (CD44, KNG1, miR-4433b-5p, miR-424-5p, and miR-199b-5p). Our study proposes potential biomarkers for MDR-TB diagnosis, and also provides a new experimental basis to understand the pathogenesis of MDR-TB.

Keywords: multidrug-resistance tuberculosis, proteomic, transcriptomic, serum, biomarkers

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How to cite this article:
Wang C, Liu CM, Wei LL, Shi LY, Pan ZF, Mao LG, Wan XC, Ping ZP, Jiang TT, Chen ZL, Li ZJ, Li JC. A Group of Novel Serum Diagnostic Biomarkers for Multidrug-Resistant Tuberculosis by iTRAQ-2D LC-MS/MS and Solexa Sequencing. Int J Biol Sci 2016; 12(2):246-256. doi:10.7150/ijbs.13805. Available from http://www.ijbs.com/v12p0246.htm