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. 2010 Aug 25:5:87-94.
doi: 10.4137/bmi.s5196.

Diagnostic markers for tuberculosis ascites: a preliminary study

Rajpal S Kashyap  1 , Sonali M Saha, Khushboo J Nagdev, Sanjeevani S Kelkar, Hemant J Purohit, Girdhar M Taori, Hatim F Daginawala
Affiliations

Diagnostic markers for tuberculosis ascites: a preliminary study

Rajpal S Kashyap et al. Biomark Insights. .

Abstract

Objective: The diagnosis of tuberculosis (TB) ascites is problematic. Delay in the diagnosis and treatment of TB ascites are considered to be major factors that contribute to the high mortality of TB. This study identifies specific protein markers in ascitic fluid which will be useful in diagnosis of TB ascites.

Methods: We used Two-Dimensional Electrophoresis, liquid chromatography-mass spectrometry/mass spectrometry, immunoblot analysis and Enzyme Linked Immunosorbent assay (ELISA) as a comprehensive quantitative proteomic screening system for the diagnosis of TB ascites.

Results: THE SCREEN IDENTIFIED SEVERAL ANTIGENS OF INTEREST: a 30-kilodalton (kDa) protein that demonstrated significant homology to the antigen 85B and 85C (Ag 85) complex; a 65-kDa protein that corresponded to Mycobacterium tuberculosis (MTB) heat shock protein 65 (65-kDa HSP), Rv0440; a 14-kDa protein and 71-kDa protein that exhibits an amino acid sequence identical to that of MTB heat shock protein 14 (14-kDa HSP), GroES; and MTB heat shock protein 71 (71-kDa HSP), Rv0350 respectively. ELISA confirmed that TB ascites patients were consistently positive for these antigens at higher rates than non-TB ascites patients.

Conclusion: The 65-kDa HSP, 71-kDa HSP, 14-kDa HSP and Ag 85 complex proteins may serve as very useful diagnostic markers for TB ascites.

Keywords: M. tuberculosis antigens; TB ascites; heat shock proteins.

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Figures

Figure 1.
Figure 1.
Two dimensional gel electrophoresis of the ‘ascitic fluid’ collected from A) TB ascites (n = 5) and B) non- TB ascites (n = 5) patients. Proteins which are up regulated in the TB ascites patients are indicated with formula image.
Figure 2.
Figure 2.
Immunoblot analysis of the identified proteins, demonstrating reactivity with the polyclonal antibodies against A) Ag 85 complex B) Mycobacterium tuberculosis heat shock protein 14 (14-kDa HSP) C) Mycobacterium tuberculosis heat shock protein 65 (65-kDa HSP) D) Mycobacterium tuberculosis heat shock protein 71 (71-kDa HSP).
See this image and copyright information in PMC

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