2012
DOI: 10.1002/pmic.201100544
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Deciphering the proteome of the in vivo diagnostic reagent “purified protein derivative” from Mycobacterium tuberculosis

Abstract: Purified protein derivative (PPD) has served as a safe and effective diagnostic reagent for 60 years and is the only broadly available material to diagnose latent tuberculosis infections. This reagent is also used as a standard control for a number of in vitro immunological assays. Nevertheless, the molecular composition and specific products that contribute to the extraordinary immunological reactivity of PPD are poorly defined. Here, a proteomic approach was applied to elucidate the gene products in the U.S.… Show more

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Cited by 52 publications
(56 citation statements)
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“…This immunodiagnostic test is based on delayed-type hypersensitivity skin reaction to tuberculin, a mixture of antigenic compounds in extracts of mycobacterial culture filtrates [19,20]. A major drawback to the TST is low specificity in certain groups of patients.…”
Section: Immunodiagnosis Of M Tuberculosis Infectionmentioning
confidence: 99%
“…This immunodiagnostic test is based on delayed-type hypersensitivity skin reaction to tuberculin, a mixture of antigenic compounds in extracts of mycobacterial culture filtrates [19,20]. A major drawback to the TST is low specificity in certain groups of patients.…”
Section: Immunodiagnosis Of M Tuberculosis Infectionmentioning
confidence: 99%
“…Twelve microliters of the tryptic peptides was analyzed with a peptide trap cartridge (Agilent, CA, USA) and the Thermo Finnigan LTQ Linear Ion Trap Mass Spectrometer (Thermo Electron, WI, USA) by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). The different parameters for mass spectrometry, such as mobile phase solutions A and B, flow rate, gradient of acetonitrile, and each survey MS scan, were set based on the information in previous studies (Borsuk et al, 2009;Cho et al, 2012). Tandem mass spectra were extracted, and the charge state was deconvoluted and deisotoped using Sorcerer 3.4 beta2 software (Sorcerer software 3.10.4, Sorcerer Web …”
mentioning
confidence: 99%
“…The protein concentration was calculated with a bicinchoninic acid (BCA) protein assay kit (Pierce, IL, USA), and 40 μg of protein was resolved on a one-dimensional 12% SDS-polyacrylamide gel. Staining, destaining, reduction, alkylation, and in-gel trypsinization were performed as previously described (Borsuk et al, 2009;Cho et al, 2012). Twelve microliters of the tryptic peptides was analyzed with a peptide trap cartridge (Agilent, CA, USA) and the Thermo Finnigan LTQ Linear Ion Trap Mass Spectrometer (Thermo Electron, WI, USA) by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS).…”
mentioning
confidence: 99%
“…However, the responses did not distinguish between animals sensitized to BCG or M. tuberculosis (15). Recently, several of the dominant proteins within PPD were defined (16). However, none of the 50 T cell antigens of PPD-S2 included the proteins in the ID93 fusion protein (Rv1813, Rv2620, Rv2608, or Rv3610).…”
Section: Discussionmentioning
confidence: 99%