Disease State Differentiation and Identification of Tuberculosis Biomarkers via Native Antigen Array Profiling

Sartain, Mark J.; Slayden, Richard A.; Singh, Krishna K.; Laal, Suman; Belisle, John T.

doi:10.1074/mcp.m600089-mcp200

Cited by 97 publications

(94 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The mycobacteria were further cultured at 37°C with agitation for 2 weeks and then washed twice with sterile phosphate-buffered saline (PBS) (Invitrogen) and inoculated in 1 liter of GAS (glycerol, alanine, salts) medium (68), a traditional Mycobacterium sp. medium used to process cells and spent filtrate for downstream proteomics applications (4,55,63). The cultures were then incubated at 37°C with agitation for 4 weeks.…”

Section: Methodsmentioning

confidence: 99%

Upregulation of the Phthiocerol Dimycocerosate Biosynthetic Pathway by Rifampin-Resistant,rpoBMutant Mycobacterium tuberculosis

Bisson

Mehaffy²,

Broeckling

et al. 2012

J Bacteriol

View full text Add to dashboard Cite

h Multidrug-resistant tuberculosis has emerged as a major threat to tuberculosis control. Phylogenetically related rifampin-resistant actinomycetes with mutations mapping to clinically dominant Mycobacterium tuberculosis mutations in the rpoB gene show upregulation of gene networks encoding secondary metabolites. We compared the expressed proteomes and metabolomes of two fully drug-susceptible clinical strains of M. tuberculosis (wild type) to those of their respective rifampin-resistant, rpoB mutant progeny strains with confirmed rifampin monoresistance following antitubercular therapy. Each of these strains was also used to infect gamma interferon-and lipopolysaccharide-activated murine J774A.1 macrophages to analyze transcriptional responses in a physiologically relevant model. Both rpoB mutants showed significant upregulation of the polyketide synthase genes ppsA-ppsE and drrA, which constitute an operon encoding multifunctional enzymes involved in the biosynthesis of phthiocerol dimycocerosate and other lipids in M. tuberculosis, but also of various secondary metabolites in related organisms, including antibiotics, such as erythromycin and rifamycins. ppsA (Rv2931), ppsB (Rv2932), and ppsC (Rv2933) were also found to be upregulated more than 10-fold in the Beijing rpoB mutant strain relative to its wild-type parent strain during infection of activated murine macrophages. In addition, metabolomics identified precursors of phthiocerol dimycocerosate, but not the intact molecule itself, in greater abundance in both rpoB mutant isolates. These data suggest that rpoB mutation in M. tuberculosis may trigger compensatory transcriptional changes in secondary metabolism genes analogous to those observed in related actinobacteria. These findings may assist in developing novel methods to diagnose and treat drug-resistant M. tuberculosis infections.A pproximately 9 million people develop active tuberculosis (TB) each year, resulting in nearly 2 million deaths annually (75). Recent progress controlling drug-susceptible TB has been made in many regions (50); however, drug resistance in Mycobacterium tuberculosis, including strains resistant to both first-line drugs, isoniazid and rifampin (multidrug-resistant [MDR] TB), has emerged as a threat to TB control worldwide (26, 65). Although international surveillance systems are inadequate, data from over 80 countries indicate that approximately 1 in 10 new cases of active TB have primary drug resistance (77), and MDR TB has been reported in essentially every country in which drug resistance has been studied (76). Treatment of MDR TB requires the use of costly and toxic second-line drugs for nearly 2 years and is associated with high rates of morbidity and mortality (65), which makes the spread of drug-resistant TB a major public health concern.Drug resistance in M. tuberculosis is due primarily to singlenucleotide polymorphisms in genes encoding key mycobacterial enzymes (6). The rpoB gene encodes the ␤-subunit of bacterial RNA polymerase, which is the target of rifampin (12...

show abstract

Section: Methodsmentioning

confidence: 99%

Upregulation of the Phthiocerol Dimycocerosate Biosynthetic Pathway by Rifampin-Resistant,rpoBMutant Mycobacterium tuberculosis

Bisson

Mehaffy²,

Broeckling

et al. 2012

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…The protein array methods are advantageous in that they require only minute amounts of patient sera (46) while enabling the simultaneous screening of large numbers of antigens in a single test (47)(48)(49)(50)(51)(52). In this methodology, purified or recombinant as well as synthetic proteins are used.…”

Section: Protein Microarraymentioning

confidence: 99%

Serologic Autoantibodies as Diagnostic Cancer Biomarkers—A Review

Zaenker

Ziman

2013

Cancer Epidemiology, Biomarkers &Amp; Prevention

130

111

View full text Add to dashboard Cite

Current diagnostic techniques used for the early detection of cancers are successful but subject to detection bias. A recent focus lies in the development of more accurate diagnostic tools. An increase in serologic autoantibody levels has been shown to precede the development of cancer disease symptoms. Therefore, autoantibody levels in patient blood serum have been proposed as diagnostic biomarkers for early-stage diagnosis of cancers. Their clinical application has, however, been hindered by low sensitivity, specificity, and low predictive value scores. These scores have been shown to improve when panels of multiple diagnostic autoantibody biomarkers are used. A five-marker biomarker panel has been shown to increase the sensitivity of prostate cancer diagnosis to 95% as compared with 12.2% for prostate-specific antigen alone. New potential biomarker panels were also discovered for lung, colon, and stomach cancer diagnosis with sensitivity of 76%, 65.4%, and 50.8%, respectively. Studies in breast and liver cancer, however, seem to favor single markers, namely a-2-HS-glycoprotein and des-g-carboxyprothrombin with sensitivities of 79% and 89% for the early detection of the cancers. The aim of this review is to discuss the relevance of autoantibodies in cancer diagnosis and to outline the current methodologies used in the detection of autoantibodies. The review concludes with a discussion of the autoantibodies currently used in the diagnosis of cancers of the prostate, breast, lung, colon, stomach, and liver. A discussion of the potential future use of autoantibodies as diagnostic cancer biomarkers is also included in this review. Cancer Epidemiol Biomarkers Prev; 22(12); 2161-81. Ó2013 AACR.

show abstract

“…Similarly, serum proteomic profiles could differentiate sputum smear-negative and smear-positive TB patients from controls with reasonable accuracy (Liu et al 2010). High-throughput screening was also applied to detect antibodies in sera against antigens from culture filtrate (Sartain et al 2006) and later to the entire Mtb proteome (Kunnath-Velayudhan et al 2010) and identified a small pool of antibodies with diagnostic potential.…”

Section: Untargeted Approaches and New Technologiesmentioning

confidence: 99%