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Demonstration of Components of Antigen 85 Complex in Cerebrospinal Fluid of Tuberculous Meningitis Patients

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Title Demonstration of Components of Antigen 85 Complex in
Cerebrospinal Fluid of Tuberculous Meningitis Patients
 
Creator Kashyap, R S
Dobos, K M
Belisle, J T
Purohit, H J
 
Subject Environmental Health
 
Description Tuberculous meningitis (TBM) is the most common form of chronic infection of the central nervous system.
Despite the magnitude of the problem, the general diagnostic outlook is discouraging. Specifically, there is no
generally accepted early confirmative diagnosis protocol available for TBM. Various Mycobacterium tuberculosis
antigens are now recognized as potential markers for diagnosis of TBM. However, their presence remains
questionable, and many of these antigens are reported in the blood but not in the cerebrospinal fluid (CSF).
This study identifies a specific protein marker in CSF which will be useful in early diagnosis of TBM. We have
demonstrated the presence of a 30-kDa protein band in CSF of 100% (n � 5) of confirmed and 90% (n � 138)
of suspected TBM patients out of 153 TBM patients. The 30-kDa band was excised from the gel, destained
extensively, and digested with trypsin. The resulting peptides were analyzed by liquid chromatography-tandem
mass spectrometry (LC-MS/MS). Partially purified proteins from CSF samples of TBM were analyzed by
two-dimensional polyacrylamide gel electrophoresis and Western blotting. Immunoblotting and enzyme-linked
immunosorbent assay (ELISA) were performed to confirm the presence of proteins in the 30-kDa protein band.
The antigen 85 (Ag 85) complex was detected in CSF of TBM patients by indirect ELISA using antibodies
against Ag 85 complex. The results of this study showed the 30-kDa protein band contained MTB proteins
Rv3804c (Ag85A) and Rv1886c (Ag 85B), both members of the Ag85 complex. This was also confirmed by using
immunotechniques such as indirect ELISA and the dot immunobinding assay. Detection of Ag85 complex was
observed in CSF of 89% (71 out of 80) of suspected TBM patients that were 30-kDa protein positive. The
observed 30-kDa protein in the CSF is comprised of the MTB Ag85 complex. This protein was earlier reported
to be present in the blood of patients with extra-central nervous system tuberculosis. Therefore, this finding
suggests that this protein can be used as a molecular marker for any type of tuberculous infection. It also
provides a more sensitive immunoassay option for the early and confirmatory diagnosis of TBM.
 
Date 2005
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://neeri.csircentral.net/81/1/Kashyap1.pdf
Kashyap, R S and Dobos, K M and Belisle, J T and Purohit, H J (2005) Demonstration of Components of Antigen 85 Complex in Cerebrospinal Fluid of Tuberculous Meningitis Patients. CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY, 12 (6). pp. 752-758.
 
Relation http://neeri.csircentral.net/81/