Proteomics of Culture Filtrate of Prevalent Mycobacterium tuberculosis Strains: 2D-PAGE Map and MALDI-TOF/MS Analysis

Kumar, Gyanendra; Shankar, Hari; Sharma, Divakar; Sharma, Prashant; Bisht, Deepa; Katoch, Vishwa Mohan; Joshi, Beenu

doi:10.1177/2472555217717639

Cited by 9 publications

(4 citation statements)

References 41 publications

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“…Expression proteomics {2-dimensional gel electrophoresis (2-DE)} coupled with MALDI-TOF-MS and bioinformatic tools has emerged as a direct approach for the separation, identification and characterization of proteins which could be used to find out potential drug targets and diagnosis of resistance as well as pathogenesis [ 11 , 12 , 13 , 14 , 15 , 16 , 17 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 ]. Since the last decade, a panel of discovery (expression), targeted proteomics and bioinformatics-based studies [ 11 , 12 , 13 , 14 , 15 , 16 , 17 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 ] have been accumulated on M. tuberculosis pathogenesis and drug resistance which showed differential expression of a panel of known and unknown proteins and suggested that these proteins might be used in diagnostics and as drug targets against drug resistant TB. We have previously reported that Rv0148 (hypothetical protein/putative short-chain type dehydrogenase/reductase) was overexpressed in aminoglycosides-resistant M. tuberculosis .…”

Section: Proteomics and Bioinformatics Explored Potential Drug Tarmentioning

confidence: 99%

Potential Alternative Strategy against Drug Resistant Tuberculosis: A Proteomics Prospect

2018

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Mycobacterium tuberculosis is one of the deadliest human pathogen of the tuberculosis diseases. Drug resistance leads to emergence of multidrug-resistant and extremely drug resistant strains of M. tuberculosis. Apart from principal targets of resistance, many explanations have been proposed for drug resistance but some resistance mechanisms are still unknown. Recently approved line probe assay (LPA) diagnostics for detecting the resistance to first and second line drugs are unable to diagnose the drug resistance in M. tuberculosis isolates which do not have the mutations in particular genes responsible for resistance. Proteomics and bioinformatic tools emerged as direct approaches for identification and characterization of novel proteins which are directly and indirectly involved in drug resistance that could be used as potential targets in future. In future, these novel targets might reveal new mechanism of resistance and can be used in diagnostics or as drug targets.

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Section: Proteomics and Bioinformatics Explored Potential Drug Tarmentioning

confidence: 99%

Potential Alternative Strategy against Drug Resistant Tuberculosis: A Proteomics Prospect

2018

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“…Ultimately, IPG strip was detached and enclose it on to second dimension electrophoresis. The second dimension was achieved on 10% gel electrophoresis for 1 h and then gels (proteins) were stained with Coomassie Brilliant Blue as described previously 15 .…”

Section: Methodsmentioning

confidence: 99%

A potent subset of Mycobacterium tuberculosis glycoproteins as relevant candidates for vaccine and therapeutic target

Yari,

Afrough,

Yari

et al. 2023

Sci Rep

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Tuberculosis (TB) remains one of the most afflictive bacterial infections globally. In high burden TB countries, surveillance, diagnosis and treatment of drug resistant TB (RR and X/MDRTB) display a crucial public health challenge. Therefore, we need new TB vaccines; diagnostic and therapeutic strategies to briskly prevent disease promotion; reduce drug-resistant TB and protect everyone from disease. The study identified various potent membrane and cell wall M. tuberculosis glycolipoproteins that are relevant for diagnostics, drug and vaccine discovery. A M. tuberculosis Proskauer and Beck broth culture was extracted for total proteins by ammonium sulfate method. After ConA-Affinity Chromatography reputed glycoproteins were collected followed by 2DE gel electrophoresis and LC Mass spectrometry. A total of 293 glycoproteins were identified using GlycoPP and IEDB database. Probable conserved trans-membrane protein (Rv0954), LpqN (Rv0583), PPE68 (Rv3873), Phosphate-binding protein (Rv0932c), PPE61 (Rv3532) and LprA (Rv1270c), had the highest glycosylation percentage value with 13.86%, 11.84%, 11.68%, 11.1%, 10.59% and10.2%, respectively. Our study discloses several dominant glycoproteins that play roles in M. tuberculosis survival, and immunogenicity. These include glycoproteins involved in antigenicity, transport and biosynthesis of M. tuberculosis cell envelope, pathogen-host interaction and drug efflux pumps, which are considered as a feasible drug targets or TB new vaccine candidates.

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“…Besides an AFB smear (Auramine O method), the homogenized sputum was used to evaluate recovery efficiency and decontamination rate (NaOH-NALC vs. PU methods). Mycobacterial culture was performed using an MGIT 960 (Becton Dickinson, Franklin Lakes, NJ, USA), and the mycobacterial strains were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS; Bruker Daltonics, Bremen, Germany) [ 17 ] or 16S rRNA gene sequencing [ 18 ].…”

Section: Methodsmentioning

confidence: 99%

Comparison of Sputum Treated with Power Ultrasound and Routine NALC-NaOH Methods for Mycobacterial Culture: A Prospective Study

Wang

Ling

et al. 2022

JCM

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Mycobacterial culture remains the gold standard for the diagnosis of active tuberculosis. However, an appropriate digestion and decontamination method is essential for the effective recovery of tubercle bacilli in culture. The study was designed to compare the efficacy of sputum treated with power ultrasound (PU) and routine NALC-NaOH methods for mycobacterial culture from clinically suspected cases of pulmonary tuberculosis. To evaluate the PU and routine NALC-NaOH methods, sputum specimens (n = 597) were studied (culturing on MGIT 960), and the performances were compared. Of the 597 samples, 89 (14.91%) sputum samples treated with the NaOH-NALC method were mycobacterial culture positive, including Mycobacterium tuberculosis (M.TB; n = 77, 12.90%) and nontuberculous mycobacteria (NTM; n = 12, 2.01%). One hundred and ten (18.43%) sputum samples treated with the PU method were culture positive, including M.TB (n = 87, 14.57%) and NTM (n = 23, 3.85%). The PU method detected 10 additional cases of M.TB and 11 additional cases of NTM when compared to the NALC-NaOH method. Statistical analysis showed that a significant difference was found in the culture-positive ratio of M.TB and NTM between the two method groups (p < 0.05). Compared with that of the NALC-NaOH method (8.04%), sputum treated with PU method (4.86%) had a significantly lower contamination rate (p < 0.05). In conclusion, our data indicate that, compared with the NALC-NaOH method, the PU method is a rapid and effective approach for mycobacterial culture when detecting active TB. However, its accurate mechanism has not been well addressed, and further investigation is still required.

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Proteomics of Culture Filtrate of Prevalent Mycobacterium tuberculosis Strains: 2D-PAGE Map and MALDI-TOF/MS Analysis

Cited by 9 publications

References 41 publications

Potential Alternative Strategy against Drug Resistant Tuberculosis: A Proteomics Prospect

Potential Alternative Strategy against Drug Resistant Tuberculosis: A Proteomics Prospect

A potent subset of Mycobacterium tuberculosis glycoproteins as relevant candidates for vaccine and therapeutic target

Comparison of Sputum Treated with Power Ultrasound and Routine NALC-NaOH Methods for Mycobacterial Culture: A Prospective Study

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