Assessment of antibody responses to antigens of Mycobacterium tuberculosis and Cysticercus cellulosae in cerebrospinal fluid of chronic meningitis patients for definitive diagnosis as TBM/NCC by passive hemagglutination and immunoblot assays

Abstract

Tanned sheep erythrocytes stabilized with pyruvic aldehyde and glutaraldehyde, called double-aldehyde-stabilized cells, were used to standardize passive hemagglutination assay (PHA) for detection of antibody responses to sonicate extract of Mycobacterium tuberculosis and Cysticercus cellulosae soluble antigens. PHA was performed in the following groups of cerebrospinal fluid (CSF) samples: group I - chronic infections of the central nervous system with the possible diagnosis of tuberculous meningitis (TBM), tuberculoma and neurocysticercosis (NCC) (n=88), and group II - controls which included (a) non-infectious non-neurological conditions (n=30), (b) infectious neurological conditions (n=21) and (c) non-infectious neurological conditions (n=133). PHA could detect anti-mycobacterial antibodies at the sensitivity level of 80.76% with a specificity of 92.4% and anti-cysticercal antibodies with a sensitivity of 100% and specificity of 92.94%. However, in 6.33% (i.e. 14/221) of group I and group II (c) CSFs both anti-mycobacterial and anti-cysticercal antibodies were detected. Immunoblot analysis of CSFs derived from TBM patients reacted predominantly to 120-kDa, 96-kDa, 65-kDa, 38-kDa, 26-kDa, 23-kDa, 19-kDa and 12-14-kDa and 4-6-kDa antigens of M. tuberculosis sonicate extract (MTSE), whilst CSFs of proven NCC reacted to >110-kDa, 96-kDa, 80-kDa, 66-68-kDa, 52-kDa and 26-28-kDa antigens of porcine whole cyst sonicate extract (PCSE). On immunoblot analysis, some of the CSFs of TBM patients were PHA positive for both MTSE and PCSE showed antibody reactivity to 70-kDa and 10-kDa antigens of C. cellulosae. Similarly CSF antibody of some Guillain Barre syndrome and myeloradiculopathy patients reacted with cysticercal antigens. But per se no cross-reactivity between MTSE and anti-cysticercal antibodies and vice-versa were observed. However, findings of this study should alert laboratory personnel especially in endemic areas to be extra careful in interpretation of antibody detection results.

A novel erythrocyte-based immunoassay for simultaneous detection of both antimycobacterial antibody response and mycobacterial antigen in human serum samples of pulmonary tuberculosis and a control group of patients using ‘a single probe’

“…The results of this study indicate that PHA had a sensitivity level of 90% with a specificity of 91% for detection of antimycobacterial antibody response (Table 1). Previous studies have shown a high sensitivity (80.76%) and specificity (92.4%) of PHA in detection of antimycobacterial antibody responses in the CSF of chronic meningitis patients (Katti, 2002). Although the detection limit of PHAI was determined to be 280 ng mL À1 (Fig.…”

Differential diagnosis of tuberculous meningitis from partially-treated pyogenic meningitis by cell ELISA

“…During the past decade, several conventional immunoassays including ELISA, dot immunobinding assays, immunoblot assays, and various molecular methods such as the polymerase chain reaction (PCR) have been reported as adjuncts in the diagnosis of TBM [4,13-15]. However, difficulties have been encountered when using many of the aforementioned techniques to differentiate TBM from PTPM.…”

Epidemiology, clinical manifestations and diagnosis of zoonotic cestode infections: an update

“…The ELISA-gp14Tcra may also be suitable for T. solium antibodies in disease-endemic populations (Bueno et al 2001;Pardini et al 2002;Peralta et al 2002). Tanned sheep red cells stabilized with pyruvic aldehyde and glutaraldehyde were used to standardize the passive hemagglutination assay (PHA) for the detection of antibody (Ab) responses to Mycobacterium tuberculosis sonicate extract and Cysticercus cellulosae soluble antigens derived from porcine whole cyst sonicate extract (Katti 2002). PHA could detect anti-mycobacterial Abs (sensitivity level: 80.7%, specificity: 92.4%) and anticysticercal Abs (sensitivity level: 100%, specificity: 92.9%) in CSF samples derived from patients with a possible diagnosis of tuberculous meningitis, tuberculoma and NCC.…”