Detection and characterisation of circulating cysticercal antigens in human cerebrospinal fluid

Katti, Muralidhar K.; Chandramukhi, A

doi:10.1016/0888-0786(90)90057-u

“…Non-speci¢c binding of human (CSF/serum) and rabbit proteins and heterophile activity of sheep erythrocytes in human CSFs were eliminated by absorbing each CSF with 10% control cells in RPHA, while in immunoblotting blotted membranes were incubated with 2% normal rabbit serum. The sensitivity limit of RPHA was 400 ng ml 31 while it was 45 ng ml 31 in our previous studies [21]. Variation in the sensitivity limit of RPHA may be due to di¡erences in titers of antibodies formed against each antigenic component in di¡erent hosts.…”

Section: Discussionmentioning

confidence: 72%

Immunodiagnosis of tuberculous meningitis: rapid detection of mycobacterial antigens in cerebrospinal fluid by reverse passive hemagglutination assay and their characterization by Western blotting

Katti¹

2001

FEMS Immunology and Medical Microbiology

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Tuberculous meningitis (TBM) is one of the commonest chronic infections of the central nervous system (CNS). Diagnosis of TBM has been a problem as it causes various clinical manifestations which can be confused with those of other chronic infections of the CNS such as neurocysticercosis (NCC), neurobrucellosis and cryptococcal meningitis, that are prevalent in many underdeveloped and developing countries. Differential diagnosis of TBM can be made by detecting circulating mycobacterial antigens in CSF by immunoassays. In this study, a reverse passive hemagglutination (RPHA) has been developed using rabbit antimycobacterial IgG for detection of circulating mycobacterial antigens in CSFs from chronic infections of the CNS in order to develop a rapid, simple, sensitive and cost-effective method. Circulating mycobacterial antigens were characterized by immunoblot assay. The sensitivity limit of RPHA was 400 ng ml(-1). RPHA was specific as antimycobacterial IgG did not show any reaction with porcine Cysticercus cellulosae which was used as a control antigen. RPHA could detect mycobacterial antigens in CSF at a sensitivity level of 94.11% with a specificity of 99.0%. Immunoblot analysis of RPHA positive CSFs revealed predominantly 30-32 kDa and 71 kDa antigens whilst 6, 86, 120, 96 and 110 kDa showed varied degree of reactivity. Antigens of masses 30-32 and 71 kDa were absent in culture filtrate of Mycobacterium tuberculosis H37Rv grown in Proskeur-Beck liquid medium. RPHA is a rapid, simple and sensitive immunological method with a long shelf life of 6-8 weeks if stabilized coated erythrocytes are stored at +4 degrees C. RPHA could be used as an additional immunodiagnostic tool in both differential diagnosis and prognosis of TBM. Immunoblot results indicate that 30-32 kDa and 71 kDa antigens are cell wall derived.

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“…Non‐specific binding of human (CSF/serum) and rabbit proteins and heterophile activity of sheep erythrocytes in human CSFs were eliminated by absorbing each CSF with 10% control cells in RPHA, while in immunoblotting blotted membranes were incubated with 2% normal rabbit serum. The sensitivity limit of RPHA was 400 ng ml −1 while it was 45 ng ml −1 in our previous studies [21]. Variation in the sensitivity limit of RPHA may be due to differences in titers of antibodies formed against each antigenic component in different hosts.…”

Section: Discussionmentioning

confidence: 67%

Immunodiagnosis of tuberculous meningitis: rapid detection of mycobacterial antigens in cerebrospinal fluid by reverse passive hemagglutination assay and their characterization by Western blotting

Katti

¹

2001

FEMS Immunology &Amp; Medical Microbiology

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Tuberculous meningitis (TBM) is one of the commonest chronic infections of the central nervous system (CNS). Diagnosis of TBM has been a problem as it causes various clinical manifestations which can be confused with those of other chronic infections of the CNS such as neurocysticercosis (NCC), neurobrucellosis and cryptococcal meningitis, that are prevalent in many underdeveloped and developing countries. Differential diagnosis of TBM can be made by detecting circulating mycobacterial antigens in CSF by immunoassays. In this study, a reverse passive hemagglutination (RPHA) has been developed using rabbit antimycobacterial IgG for detection of circulating mycobacterial antigens in CSFs from chronic infections of the CNS in order to develop a rapid, simple, sensitive and cost-effective method. Circulating mycobacterial antigens were characterized by immunoblot assay. The sensitivity limit of RPHA was 400 ng ml(-1). RPHA was specific as antimycobacterial IgG did not show any reaction with porcine Cysticercus cellulosae which was used as a control antigen. RPHA could detect mycobacterial antigens in CSF at a sensitivity level of 94.11% with a specificity of 99.0%. Immunoblot analysis of RPHA positive CSFs revealed predominantly 30-32 kDa and 71 kDa antigens whilst 6, 86, 120, 96 and 110 kDa showed varied degree of reactivity. Antigens of masses 30-32 and 71 kDa were absent in culture filtrate of Mycobacterium tuberculosis H37Rv grown in Proskeur-Beck liquid medium. RPHA is a rapid, simple and sensitive immunological method with a long shelf life of 6-8 weeks if stabilized coated erythrocytes are stored at +4 degrees C. RPHA could be used as an additional immunodiagnostic tool in both differential diagnosis and prognosis of TBM. Immunoblot results indicate that 30-32 kDa and 71 kDa antigens are cell wall derived.

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“…The relative merits of antigen detection versus antibody analysis have been reviewed elsewhere [7]. My colleagues and I selectively demonstrated 2 cysticercal antigens in human CSF from patients with proven NCC, with molecular masses of 24-28 and 64-68 kDa, of which the 24-28 kDa antigen was found to be highly specific [8].…”

Section: Assessment Of Specificity Of a Recombinant 10-kda Protein Anmentioning

confidence: 99%

Assessment of Specificity of a Recombinant 10‐kDa Protein Antigen in Differential Diagnosis of Neurocysticercosis

Katti

¹

2000

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mydia pneumoniae stimulate cytokine production in human blood mononuclear cells. Eur J Immunol 2000; 30:541-9 . 4. Kol A, Sukhova G, Lichtman A, Libby P. Chlamydial heat shock protein 60 localizes in human atheroma and regulates macrophage tumor necrosis factor-alpha and matrix metalloproteinase expression. Circulation 1998; 98:300-7. 5. Kol A, Bourcier T, Lichtman AH, Libby P. Chlamydial and human heat shock protein 60s activate human vascular endothelium, smooth muscle cells, and macrophages. J Clin Invest 1999; 103:571-7. 6. Kalayoglu MV, Byrne GI. Induction of macrophage foam cell formation by Chlamydia pneumoniae. J Infect Dis 1998; 177:725-9. 7. Kalayoglu MV, Byrne GI. A Chlamydia pneumoniae component that induces macrophage foam cell formation is chlamydial lipopolysaccharide. Infect Immun 1998; 66:5067-72. 8. Kalayoglu MV, Miranpuri GS, Golenbock DT, Byrne GI. Characterization of low density lipoprotein uptake by murine macrophages exposed to Chlamydia pneumoniae. Microbes Infect 1999; 6:409-18. 9. Leeuwenburgh C, Rasmussen JE, Hsu FF, Mueller DM, Pennathur S, Heinecke JW. Mass spectrometric quantification of markers for protein oxidation by tyrosyl radical, copper, and hydroxyl radical in low density lipoprotein isolated from human atherosclerotic plaques.

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Detection and characterisation of circulating cysticercal antigens in human cerebrospinal fluid

Cited by 10 publications

References 12 publications

Immunodiagnosis of tuberculous meningitis: rapid detection of mycobacterial antigens in cerebrospinal fluid by reverse passive hemagglutination assay and their characterization by Western blotting

Immunodiagnosis of tuberculous meningitis: rapid detection of mycobacterial antigens in cerebrospinal fluid by reverse passive hemagglutination assay and their characterization by Western blotting

Immunodiagnosis of tuberculous meningitis: rapid detection of mycobacterial antigens in cerebrospinal fluid by reverse passive hemagglutination assay and their characterization by Western blotting

Assessment of Specificity of a Recombinant 10‐kDa Protein Antigen in Differential Diagnosis of Neurocysticercosis

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