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

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.

Identification of a plasma microRNA profile in untreated pulmonary tuberculosis patients that is modulated by anti-mycobacterial therapy

“…Other recent biomarker studies have identified additional miRNA that may have biomarker potential to aid TB diagnosis. miRs-144, -223, -29a, -155 and 424 have all been reported as providing AUC over 0.8 or sensitivity and specificity over 80% as a single miRNA biomarker aiding TB diagnosis (12,(22)(23)(24)(25). Of these, only miRs -223 and -29a were significantly regulated in our exploratory study, with only miR-29a remaining significantly regulated in the validation study.…”

“…(9,10). Studies have shown changes in blood miRNA levels in small numbers of patients with active TB that have biomarker potential (11)(12)(13).…”

Analysis of the plasma proteome using iTRAQ and TMT‐based Isobaric labeling

“…Rheumatoid arthritis (RA): Cheng et al, 140 Dwivedi et al, 141 Ortea et al, 142 Serada and Naka, 143 135 Wang et al, 136 Xu et al, 137 Considering the influence of cerebrovascular irregularities, the plasma proteomics of acute ischemic stroke 123 has been studied. Also regarding a subtype of the latter, that is, lacunar infraction, Datta et al enriched mircovesicles from plasma to determine whether differences in their proteomic profiles could be used to predict adverse outcome in the surviving patients.…”

“…134 Although cord blood serum can be richer in intracellular proteins and more varied than matured serum, the study was conducted with older MS instrumentation (Applied Biosystems, QSTAR-Pulsar) and they reported quantification of in the order of 200 proteins. Other studies of respiratory afflictions have included tuberculosis [135][136][137][138] and Mycoplasma pneumoniae (MP) infections 139 (see below). Examples of these applications are indicated in Table 5A.…”

“…Isobaric labeling-based studies of the plasma proteome in response to infection have ranged from bacterial, parasitic (ie, Malaria 170 ) and viral encounters (see Table 5A). Bacterial studies include iTRAQ studies of pulmonary tuberculosis (PTB) in humans [135][136][137][138] and mycobacterial infections in cattle. 45 In the works of Xu et al and Wang et al, differences in serum proteome were studied in association with PTB.…”