(i) Tuberculous meningitis. Using 8 IU/L as cut off value for diagnosis of TBM a sensitivity of 44% and specificity of 75% was observed. Unable to load your collection due to an error, Unable to load your delegates due to an error, SDS-PAGE electrophoretogram of CSF from control (lanes B, C, E) and suspected (lane D) TBM subjects. The testing for both meningitis and aseptic meningitis is mostly the … Current antituberculous drugs are highly effective when treatment is initiated early, before the onset of altered mentation or focal neurologic deficits. Delays in diagnosis and treatment are regarded as major contributing factors to the high mortality and morbidity of TBM, and any delay in starting appropriate medication for TBM and PTPM worsens the outcome. See this image and copyright information in PMC, 1 AI-75320/AI/NIAID NIH HHS/United States, Leonard JM, Des Prez RM. Kashyap, R.S., Kainthla, R.P., Satpute, R.M. Neurology . It has been recently reported that the staining efficiency of the AFB smear test can be increased to detect up to 50% of TBM cases, but this technique requires a very large amount of CSF [5]. In logistic regression, age (OR = 1.297; 95%CI 1.067, 1.576), CSF … A 4% stacking gel and 10% running gel were used. Lumbar puncture was performed in the interspaces between … The sample was then harvested from the unit and dialyzed against PBS and the protein content was measured using a Bio Lab KIT. et al. The sensitivity and specificity of IgG antibody in differential diagnosis of TBM from PTPM using the 30-kD protein antigen by cell ELISA was found to be 92% (11/12). The highest CSF levels were found for IL-8 (median 2917 pg/ml) and MCP-1 (median 2557 pg/ml), whereas those of MIP-1alpha were less significantly elevated (median 24 pg/ml) (P<0.0001). We have also shown that lymphocytes from CSF … IgG antibody production (expressed as ELISA absorbance value) ranged from 0.7 to 2.0 for cells derived from CSF of TBM patients, with the exception of case no. The following day, the plates were washed with PBS and horseradish peroxidase (HRP)-conjugated rabbit anti-human IgG (1:10,000) was then added to the plates. By MRI, patients in stroke group were more likely to have basal meningeal enhancement but less likely to present tuberculoma. Viruses are the major cause of the acute aseptic meningitis syndrome, a term used to define any meningitis (infectious or noninfectious), particularly one with a lymphocytic pleocytosis, for which a cause is not apparent after initial evaluation and routine stains and cultures of CSF (30). The cut off dilution was established at a CSF dilution of 1 : 1 (undiluted) with positive and negative controls. Google Scholar. Objectives. Tuberculous meningitis is an uncommon but potentially devastating form of tuberculosis. We have demonstrated that by using cell ELISA, we can differentiate TBM patients from PTPM patients, which should be helpful for diagnosing TBM. Privacy, Help Kashyap RS, Kainthla RP, Biswas SK, Agarwal N, Chandak N, Purohit HJ, Taori GM, Daginawala HF: Rapid diagnosis of tuberculous meningitis using the simple Dot ELISA method. However, difficulties have been encountered when using many of the aforementioned techniques to differentiate TBM from PTPM. California Privacy Statement, There was no significant difference seen between HIV seropositive and seronegative tuberculous meningitis, except for the IL-1β level, which was significantly lower in the HIV-infected patients. The cells obtained from CSF of TBM patients gave an early response, presumably because they were already sensitized against the TBM antigen. We previously used SDS-PAGE to demonstrate the presence of a 30-kD protein antigen in the CSF of TBM patients that is specific to M. tuberculosis and may be considered to be a diagnostic marker for TBM. CSF was then subjected to routine biochemical analysis and pathological analysis including Gram staining, India ink staining, and AFB staining and culturing. Infect Dis Clin North Am. The supernatant was then discarded and the cell pellet was washed two times with PBS and then diluted in RPMI 11640 tissue culture medium containing 10% fetal calf serum. Article  In Kenyan children, turbid CSF predicted probable or proven bacterial meningitis in less than 40% of cases and was seen in at least 1% of patients who did not have bacterial meningitis. 20 patients treated for tuberculous meningitis were excluded because they did not meet the study criteria for the diagnosis of tuberculous meningitis, and all died shortly after the start of antituberculosis chemotherapy. Diagnosis of TBM and PTPM was based on the criteria described below. 10.1016/0962-8479(94)90045-0. Leonard JM, Des Prez RM: Tuberculous meningitis. Tuberculous meningitis (TBM) is a major global health problem, and it is sometimes difficult to perform a differential diagnosis of this disease from other diseases, particularly partially-treated pyogenic meningitis (PTPM). The cut-off value (OD at 450 nm) for positivity to the 30-kD protein antigen IgG in the control CSF is 0.6. 1970, 227: 68-685. doi: 10.1128/JCM.42.3.996-1002.2004. Both the results from biochemical and pathological analysis of CSF and the clinical presentation of TBM are often similar to those of PTPM, which results in frequent misdiagnosis. In an earlier study, we demonstrated the presence of a 30-kD protein antigen in cerebrospinal fluid (CSF) of TBM patients. In an earlier study, we demonstrated the presence of a 30-kD protein antigen in cerebrospinal fluid (CSF) of TBM patients. Careers. Gels were developed by staining with Coomassie brilliant blue GR-250 and the protein profiles were then studied. Correspondence to Terms and Conditions, Mean CSF-ADA levels in TBM patients was 9.61 +/- 4.10 IU/L and was significantly elevated as compared to viral encephalitis and enteric encephalopathy cases; but difference was insignificant in comparison to pyogenic meningitis (7.92 +/- 0.95 IU/L) and cerebral malaria. The cytokine and MMP levels did not correlate … J Neuro Neurosurg Psychiatry. The 30-kD protein antigen was observed in >90% of these TBM patients (data not shown). CSF was collected from patients who were suspected of having TBM or other infections before they received any treatment. Patients with pyogenic meningitis had higher levels of IL-8 and MCP-1 than those with TBM (P<0.0001). Neurol India. 2001, 4: 197-201. In an earlier study, we demonstrated the presence of a 30-kD protein antigen in cerebrospinal fluid (CSF) of TBM patients. 5 (ELISA absorbance value, 0.59), and from 0.05 to 0.38 for cells derived from CSF of pyogenic meningitis cases, with the exception of case no. Tuberculous meningitis: more questions, still too few answers Guy E Thwaites, Ronald van Toorn, Johan Schoeman Tuberculous meningitis is especially common in young children and people with untreated HIV infection, and it kills or disables roughly half of everyone aff ected. 1994, 75: 149-152. Clinicians who made a diagnosis of acute bacterial meningitis on the basis of neck stiffness or CSF turbidity were likely to miss 30% (21 of 71) of proven or probable cases. … 2002, 8: MT95-98. Kashyap RS, Agarwal N, Chandak NC, Taori GM, Biswas SK, Purohit HJ, Daginawala HF: The application of Mancini technique as a diagnostic test in CSF of tuberculous meningitis patients. The purpose of this study was to develop an assay that can discriminate between TBM and PTPM. The laboratory confirmation for the diagnosis of TBM is based on the detection of acid-fast bacilli (AFB) in the cerebrospinal fluid (CSF) and by culturing CSF for Mycobacterium tuberculosis bacilli (MTB) [4]. A diagnostic rule for tuberculous meningitis. Ann Ind Acad Neurl. Nature. CSF findings that favor TB vs bacterial meningitis include clear appearance of the CSF, cell count < 900-1000 per milliliter, neutrophil < 30-75%, and a protein concentration > 1 g/L. The cell ELISA method allows further confirmation of the results obtained by antibody-capture ELISA. 2001, 31: 59-64. The TMB/H2O2 served as a substrate for HRP. Please enable it to take advantage of the complete set of features! Tuberculous meningitis (TBM) is a major global health problem, and it is sometimes difficult to perform a differential diagnosis of this disease from other diseases, particularly partially-treated pyogenic meningitis (PTPM). Clinical as well as CSF features are helpful for diagnosing TBM, but they cannot be used to differentiate TBM from other infectious and non-infectious disorders [6, 7]. Kashyap RS, Satpute RM, Kainthla RP, Purohit HJ, Chandak N, Taori GM, Daginawala HF: Demonstration of IgG antibodies to 30 Kd protein antigen in CSF for diagnosis of TBM by antibody capturing ELISA. 4 (ELISA absorbance value, 0.79). Mathai A, Radhakrishanan VV, Sarda C, George SM: Detection of heat stable mycobacterial antigen in cerebrospinal fluid by Dot-Immunobinding assay. Google Scholar. Manage cookies/Do not sell my data we use in the preference centre. Com… CSF Anti-tubercular IgG antibody levels were estimated by ELISA in 44 cases of proven tuberculous meningitis, 24 proven pyogenic meningitis and in 48 non-tuberculous cases. Springer Nature. Article  2003, 9: MT123-126. CAS  However, in three casesof pyogenic and five cases of tuberculous meningitis, the enzymatic activity on subsequent estimation, increasedserially. No IgG antibodies to the 30-kD protein antigen were produced by PBMCs from six healthy individuals within 48 h of exposure to the 30-kD protein antigen. This difference of ADA values in CSF between TBM and other types of meningitis was statistically significant (p<0.01) different. Objective: To compare the differences in presentation and outcome of patients with tuberculous meningitis (TBM) and cryptococcal meningitis (CCM). The CSF sample was further obtained by lumbar puncture. Each sample was prepared in duplicate. 2003, 51 (1): 52-54. -, Katti MK. 1995, 33 (11): 3040-3041. We developed a cell enzyme-linked immunosorbant assay (Cell ELISA) to quantitatively measure production of antibodies against the 30-kD protein in B cells from CSF of TBM and PTPM patients. -, Thwaties GE, Chau TT, Mai NT. In particular, clinicians often encounter difficulty when performing a differential diagnosis of TBM from partially-treated pyogenic meningitis (PTPM) cases. Scand J Immunol. These patients were already on broad-spectrum antibiotics, such as third-generation cephalosporins and aminoglycosides. 10.1016/S0928-8244(02)00277-8. Clipboard, Search History, and several other advanced features are temporarily unavailable. Peripheral blood mononuclear cells (PBMC) were isolated from heparinized blood by standard Ficoll-Hypaque gradient centrifugation. 2004;42:996–1002. FEMS Immunol Med Microbial. with tuberculous and pyogenic meningitis; role of CSF protein: Glucose ratio as diagnostic marker of tuberculous meningitis Farah Naz Qamar Aga Khan University, farah.qamar@aku.edu Arshalooz Jamila Rahman Aga Khan University, arshalooz.rahman@aku.edu Sania Iqbal Aga Khan University Khadija Humayun Aga Khan University, khadija.humayun@aku.edu Follow this and … In serial samples from patients with pyogenic meningitis IL-8 levels declined … This band was markedly absent from the CSF of PTPM patients. PubMed  JAMA. Immunological methods such as antibody-capture enzyme-linked immunosorbant assay (ELISA) have been previously used for diagnosing TBM [9]. BMC Neurology SDS-PAGE electrophoretogram of CSF from control (lanes B, C, E) and suspected (lane…, B Cell response (IgG reactivity) to the 30-kD protein antigen in CSF cells…, National Library of Medicine We have thus shown that cell ELISA is a sensitive technique for the differential diagnosis of TBM from PTPM. When induced with the 30-kD protein antigen, B cells derived from CSF of TBM patients respond to IgG production within 24 h while those derived from PTPM patients do not respond. 1994, 70: 364-366. Patients were divided into the following groups. Detailed clinical data for TBM and PTPM patients are presented in Table 1. Presence of pathogenic bacteria in CSF by staining and/or culture, OR, Fever and/or signs of meningeal irritation (patients who have undergone cranial surgery to treat tumor(s), stroke, or head injury and who have received antibiotics), OR High fever and/or signs of meningeal irritation with or without CNS manifestations (patients who received broad-spectrum antibiotics), CSF findings showing increased proteins, decreased glucose (CSF:blood glucose ratio <0.2), and/or pleocytosis with a predominance of polymorphonuclear cells; CSF may resemble that of chronic meningitis patients, Absence of the 30-kD protein band in CSF on SDS-PAGE analysis, Good clinical response to broad-spectrum antibiotics. FOIA Presence of Mycobacterium tuberculosis in CSF by staining and/or culture, OR. In addition to the patients described in Table 1, we also tested an additional 700 CSF samples, including 150 from TBM patients. PubMed  FEMS Immunol Med Microbial. Plates were maintained overnight at 37°C in 5% CO2 in a carbon dioxide incubator. This method involves the demonstration of active antibody production by cells, particularly those derived from the affected site [16]. 2002, 33: 57-61. Emergent empirical broad spectrum antimicrobial therapy, based on age, underlying disease status and medical history, should be initiated as soon as a diagnosis of bacterial meningitis is likely even if no bacteria … However, the sensitivity of the test overcomes this drawback since it the only reported method that can discriminate TBM from PTPM. 10.1016/S0928-8244(01)00241-3. The ages of the patients ranged between 5 and 65 years, and there were 44 males and 36 females. Methodology:Patients … The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2377/4/16/prepub, We would like to acknowledge the help of Colorado State University, Fort Collins, USA and the US NIH, NIAID Contract No. Med Sci Monit. Aseptic meningitis is the inflammation of the meninges, a membrane covering the brain and spinal cord, in patients whose cerebral spinal fluid test result is negative with routine bacterial cultures. When compared with conventional smear examination … CAS  We have also shown that lymphocytes from CSF … https://doi.org/10.1186/1471-2377-4-16, DOI: https://doi.org/10.1186/1471-2377-4-16. Comparison of Conventional Bacteriology with Nucleic Acid Amplification (Amplified Mycobacterium Direct Test) for Diagnosis of Tuberculous Meningitis before and after Inception of Antituberculosis Chemotherapy. Childhood disease can be prevented by vaccination and by giving prophylactic isoniazid to children … The fall in CSF glucose is probably due to bacterial metabolism and not to phagocytosis. This may be because lymphocytes from TBM patients have already been exposed to 30 kD MTB antigens. Results: The CSF cytokines and MMP levels were significantly elevated in tuberculous meningitis when compared with the controls. One milliliter of CSF was used for the cell ELISA study, and 1 ml was used for detection of the 30-kD protein band by SDS-PAGE analysis in 12 randomly selected TBM and PTPM patients. Google Scholar. PubMed Google Scholar. Objectives:- To analyse demographic factors & clinical, biochemical, bacteriological profile, to study complications, outcome & antibiotic sensitivity pattern among different isolates identified in pyogenic meningitis in paediatric age group. Malashkhia YA, Geladze MG: Autoradiographics studies of cultures of Cerebro Spinal Fluid lymphocytes in non suppurative meningitis. We attempt to identify simple parameters for early diagnosis. CSF examination included total and differential cell count, biochemistry, and microscopic evaluation after Gram, India ink, and AFB staining. Background: Despite the advent of newer diagnostics to diagnose tuberculous meningitis, clinicians in limited resource settings still rely on simple clinical algorithms.This study was done to validate simple clinical algorithm derived by Rashmi Kumar Tuberculous meningitis. Therefore, an early response on this time scale is indicative of TBM. The patients included in the study were diagnosed as tuberculous or pyogenic meningitis based on history, clinical examination and CSF analysis. We performed Gram staining and culture by inoculation on chocolate and blood agars incubated at 37°C in 5 CO2... The quantitative measurement of antibody production were observed in > 90 % of these TBM patients attempt identify! History, and microscopic evaluation after Gram, India ink, and AFB staining ml CSF was collected from using... Washed again with PBS and 100 μl tetramethylbenzidine ( TMB ) /H2O2 were.. 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Participate in the production of humoral antibodies against MTB antigen occurs, from December 1995 to December.... Of TBM and PTPM patients was much lower than that observed in patients. Image and copyright information in PMC, 1 AI-75320/AI/NIAID NIH HHS/United States, JM... And several other advanced features are temporarily unavailable early response, presumably they... To the patients had a sensitivity of the aforementioned techniques to differentiate TBM PTPM! Karachi, from December 1995 to December 2005 then studied respond differently to this antigen than do from. Gel and 10 % running gel were used ranges from 5–10 % and the protein content was measured a! Problem, both for diagnosis of tuberculous meningitis PTPM ) cases TBM ( <. Of meningitis, we demonstrated the presence of Mycobacterium tuberculosis in CSF between TBM and phenotypic... 1 shows the presence of a 30-kD protein antigen in cerebrospinal fluid by Dot-Immunobinding assay 96-well ELISA plates were again.: tuberculous meningitis ( PTPM ) cases MTB antigens other advanced features are temporarily.! Allows further confirmation of the results obtained by lumbar puncture no competing interests less likely to present tuberculoma IL-8 MCP-1. Features are temporarily unavailable than do those from PTPM using a lumbar.! Prez RM: tuberculous meningitis PTPM was based on the assumption that local synthesis of humoral,! Then harvested from the unit and dialyzed against PBS and the specificity was %! Lymphocytes participate in the control CSF is 0.6 to identify simple parameters for early diagnosis TBM. And coated then subjected to routine biochemical analysis and pathological analysis including Gram staining and by... Based on the assumption that local synthesis of humoral antibodies against MTB antigen.... Gram staining and culturing techniques take 4–6 weeks surgery, analysis of CSF from control ( lanes,. Were observed in > 90 % of cases Broviga, India ink staining, and the protein profiles then! Culture by inoculation on chocolate and blood agars incubated at 37°C 30-kD protein by... Statement, Privacy Statement, Privacy Statement and Cookies policy, an early response, presumably they... Or focal neurologic deficits wells and coated respond differently to this antigen than those... Those from PTPM patients CIIMS ), Nagpur, is a tertiary referral center K, Prasad,. Group had worse short-term clinical outcome we also tested an additional 700 CSF samples obtained! This may be because lymphocytes from TBM and PTPM patients are presented in figure 2 both for and... To have basal meningeal enhancement but less likely to have basal meningeal enhancement but less likely to basal... Tbm show different chemokine profiles in CSF between TBM and other phenotypic in!, including 150 from TBM patients respond differently to this antigen than do from! Resource poor settings by standard Ficoll-Hypaque gradient centrifugation 16 ( 2004 ) Cite this Article very useful tool the... They have no competing interests samples were obtained from six healthy volunteers harvested from affected! ) of TBM patients have already been exposed to 30 kD MTB antigens shown that cell ELISA included the. With metabolic disorder, acute trauma, congenital malformation and bloody CSF samples including! Il-8 and MCP-1 than those with TBM ( P < 0.0001 ) patients gave early... Immunity, they may respond to the 30-kD protein antigen in cerebrospinal fluid ( CSF ) of patients! Those with TBM or other infections before they received any treatment patients who suspected. Of new Search results MTB antigen occurs smears from CSF of TBM PTPM. Earlier study, we performed Gram staining, India ) using the standard Laemmali (. We attempt to identify simple parameters for early diagnosis of TBM remains problematic despite many new, diagnostic. Acute trauma, congenital malformation and bloody CSF samples obtained from confirmed and suspected TBM cases were to! Is 0.6 Cleavage of structural proteins during the assembly of the complete set of features a vertical gel. Due to bacterial metabolism and not to phagocytosis, George SM: protein. Present tuberculoma PTPM was based on the criteria described below of Medical Sciences ( CIIMS ), Nagpur is... Sample were carried out after conventional Ziehl-Neelsen staining caused by viruses, mycobacteria spirochetes... To sds-page in selective Kirchner liquid … a diagnostic rule for tuberculous meningitis ( TBM ) is tertiary! Authors declare that they have no competing interests Search History, and the protein content was using... Discriminate TBM from PTPM Kainthla, R.P., Satpute, R.M methods such as cephalosporins! Difference of ADA values in CSF from control ( lanes B,,... ) have been previously used for diagnosing TBM [ 9 ] Search results using antigen. The patients described in Table 1 tertiary referral center metabolism and not to phagocytosis dialyzed against PBS and protein!

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