Urinary proteomic analysis to identify a potential protein biomarker panel for the diagnosis of tuberculosis

Abstract

Tuberculosis (TB) is caused by Mycobacterium tuberculosis and is one of the primary causes of death worldwide. Rapid and accurate diagnosis of TB is one of the most direct means to reduce the incidence of TB. In this study, urinary proteomic profiling of TB patients and non-TB individual controls (HCs) was performed, and differentially expressed urinary proteins between TB and HCs were compared and exclusively expressed proteins in TB patients were selected to establish a clinically useful disease marker panel. In total, these top 11 targeted proteins with 265 peptides were scheduled for multiple reaction monitoring validation analysis by using urine samples from 52 TB patients and 52 HCs. The result demonstrated that a three-protein combination out of the five-protein panel (namely P22352, Q9P121, P15151, Q13291, and Q8NDA2) exhibited sensitivity rate of 82.7% in the diagnosis of TB. Furthermore, the three-protein combination could differentiate TB from the latent tuberculosis (LTB) effectively, which exhibited specificity rate of 92.3% for the diagnosis of TB from the LTB category. Although more numbers of clinical samples are required for further verification, the results provided preliminary evidence that

A Peptidomic Approach to Identify Novel Antigen Biomarkers for the Diagnosis of Tuberculosis

“…All the peptide mixtures were concentrated using a Speed-vac centrifuge (Eppendorf, Germany), dissolved in 0.1% formic acid, and analyzed in an ultra-performance liquid chromatograph (UPLC) system (Waters, Milford, MA) coupled with an LTQ Orbitrap Velos mass spectrometer (ThermoFisher Scientific, Waltham, MA) as previously reported. 26 , 27 Proteome Discoverer software (version 2.4.0.305, Thermo Fisher Scientific, Waltham, MA) were used to process the raw data with search algorithms SEQUEST (version 1.3, Thermo Fisher Scientific, Waltham, MA) against Mycobacterium tuberculosis (strain ATCC 25618/H37Rv) database from Swiss-Prot database (release 2021_04). All the serum samples from the same condition were pooled and the LC-MS/MS analysis was performed in three technical replicates.…”

“…All the peptide mixtures were concentrated using a Speed-vac centrifuge (Eppendorf, Germany), dissolved in 0.1% formic acid, and analyzed in an ultra-performance liquid chromatograph (UPLC) system (Waters, Milford, MA) coupled with an LTQ Orbitrap Velos mass spectrometer (ThermoFisher Scientific, Waltham, MA) as previously reported. 26,27 Proteome Discoverer software (version 2.4.0.305, Thermo Fisher Scientific, Waltham, MA) were used to process the raw…”

From simple to complex: Protein‐based biomarker discovery in tuberculosis

“…Mass spectrometry [36,37], [38] # Other ELISA [40,41], [130,131] Mass spectrometry [42] # multi-omics.…”

Challenges and the Way forward in Diagnosis and Treatment of Tuberculosis Infection

“…Selected targets were validated using multiple reaction monitoring (MRM), and ROC analysis showed that a combination of five biomarkers, i.e., P22352 (glutathione peroxidase 3), Q9P121 (neurotrimin), P15151 (poliovirus receptor), Q13291 (signaling lymphocytic activation molecule family 1), and Q8NDA2 (hemicentin-2), had the best accuracy in the diagnosis of ATB. Out of these five panels, a three-protein combination (Q9P121, P15151, and Q8NDA2) showed a sensitivity rate of 82.7% in the diagnosis of ATB from non-TB and a specificity of 92.3% for the diagnosis of ATB from the TBI group [ 202 ].…”