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Korean J Parasitol > Volume 30(4):1992 > Article

Original Article
Korean J Parasitol. 1992 Dec;30(4):299-307. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1992.30.4.299
Copyright © 1992 by The Korean Society for Parasitology
Measurement of 150 kDa protein of Taenia solium metacestodes by antibody-sandwich ELISA in cerebrospinal fluid of neurocysticercosis patients
S Y Cho,Y Kong,S I Kim,** and S Y Kang
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea.
Abstract

An antigenic protein in cystic fluid of Taenia solium metacestodes (CF) of 150 kDa was measured by antibody-sandwich ELISA in serum and cerebrospinal fluid (CSF) of neurocysticercosis patients. Capture antibodies were rabbit antisera against CF (RACF) and a monoclonal antibody (MAb) against 150 kDa protein in CF. Lower limit of antibody-sandwich ELISA was 8 ng/ml of the protein. Except CF, no tested helminths extracts reacted. Levels of the protein in 351 sera from 255 patients (55 surgery confirmed and 202 antibody and CT/MRI confirmed) were below sensitivity of the assay. Of 276 CSF from 212 patients, 31 samples (11.2%) showed positive findings. This assay, therefore, was not sensitive enough to be a diagnostic. Instead, the 150 kDa protein appeared in CSF in such situations as in 2 days after praziquantel treatment, or as in a patient infected with a racemose cysticercus with degenerated cyst wall. Of cases whose follow-up CSF were assayed, 2 cases showed that the protein appeared intermittently. These results suggest strongly that appearance of free 150 kDa protein is associated with cyst wall rupture. In CSF which contained the 150 kDa protein over 61 ng/ml, the protein was recognized in SDS-PAGE before and after immunoprecipitation.

Figures


Fig. 1
Relation between 150 kDa protein concentration in Cf and absorbance by antibody-sandwich ELISA. The concentration was calculated as 70% of whole proteins in CF (6.0 mg/ml) as measured by densitometry (Cho et al., 1988). The standard curve was linear in protein range of 8~60 ng/ml. This range was considered to be the best measurement interval form the 150 kDa protein.


Fig. 2
SDS-PAGE findings of CSF and surgically collected cystic fluid of NCC patients. The samples were electrophoresed on 10~15% reducing gel with the supply of 30 mA constant current. Mr: Molecular mass in kDa, Lanes 1 and 5 : Surgically collected cystic fluid, Lanes 2,3,4,6,7,8,9, and 10: NCC patients CSF, CF: Cystic fluid collected from a pig, C: Band C protein (150 kDa protein).


Fig. 3
SDS-PAGE and immunoblot findings of CSF from NCC patients after the immunoprecipitation. The proteins were separate using 10~17.5% pradient gel. Mr: Molecular mass in kDa, N: CSF from other mcurologic patient, 1~7: CSF collected each NCC patient, CF: Cystic fluid collected from a pig. (A): MAb bound antigenic subunits of molecular mass of 10 kDa were recognized at the bottom of each lane (arrow head). (B): When this immune complex was immunoblotted with RACF, 10 kDa bands of 150 kDa protein were reacted (arrow head). Heavy and light chains of rabbit IgG (in Pansorbin) were non-specifically reacted with RACF.

Tables


Table 1
Number of NCC cases and their samples assayed in this study


Table 2
Range of 150 kDa protein concentration (ng/ml) in CSF of NCC patients


Table 3
Antibody and 150 kDa protein levels in serum and CSF of a NCC patient who was treated with proziquantel since Now. 11, 1987


Table 4
Antibody and 150 kDa protein levels in serum and CSF of a NCC patient with hydrocephalus


Table 5
Antibody and 150 kDa protein levels in serum and CSF of another NCC patient with hydrocephalus and multiple low densities

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