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A sensitive ELISA for glial fibrillary acidic protein: application in CSF of children
L E Rosengren1, G Ahlsén, M Belfrage
1Institute of Neurobiology, University of Göteborg, Sweden.
Insights
A new ELISA method accurately measures glial fibrillary acidic protein (GFAP) in cerebrospinal fluid (CSF). Elevated GFAP levels in children with autism suggest reactive astrogliosis, aiding in diagnosing brain disorders.
Area of Science:
- Neuroscience
- Biochemistry
- Immunology
Background:
- Glial fibrillary acidic protein (GFAP) is a key marker of astrocytes in the central nervous system (CNS).
- Elevated GFAP levels in cerebrospinal fluid (CSF) can indicate CNS damage or disorders.
- Accurate and sensitive measurement of GFAP is crucial for diagnosing neurological conditions in children.
Purpose of the Study:
- To develop and validate a sensitive enzyme-linked immunosorbent assay (ELISA) for quantifying glial fibrillary acidic protein (GFAP).
- To investigate GFAP and S-100 protein levels in the CSF of normal children and children with infantile autism.
- To assess the utility of GFAP and S-100 protein measurements in differentiating pediatric brain disorders.
Main Methods:
- A novel sandwich ELISA utilizing the biotin-avidin system was developed for GFAP determination with a sensitivity of 16 pg/ml.
- GFAP and S-100 protein concentrations were measured in CSF samples from normal children and children with autism.
- Commercial ELISA kits were used for S-100 protein measurements.
Main Results:
- The developed GFAP ELISA demonstrated high sensitivity and precision.
- CSF GFAP levels in normal children correlated with age, with a more pronounced increase than S-100 protein.
- Children with infantile autism exhibited higher CSF GFAP levels compared to age-matched controls, while S-100 protein levels were similar.
Conclusions:
- The sensitive GFAP ELISA enables accurate measurement of low GFAP concentrations in pediatric CSF.
- Elevated GFAP with normal S-100 protein in CSF suggests reactive astrogliosis in the CNS.
- Combined GFAP and S-100 protein assays can help distinguish between acute and chronic pediatric brain disorders.
Abstract:
In the present study we describe a sensitive ELISA for determination of glial fibrillary acidic protein (GFAP). To validate the method combined determinations of GFAP and S-100 protein were performed in cerebrospinal fluid (CSF) of normal children and children with autism. The GFAP ELISA is of sandwich type and uses the biotin-avidin system. Sensitivity was 16 pg/ml. Between-day precision was 0.079 (coeff. of variance). S-100 protein concentrations were measured using a commercially available ELISA kit. Normal CSF from children and young adults were analysed. The CSF levels of GFAP in normal children were low (16-163 pg/ml). Both GFAP and S-100 protein concentrations correlated with age (P < 0.01 and P < 0.05, respectively), but the GFAP increment was more pronounced, probably reflecting the age-dependent expansion of the fibrillary astrocytes in the central nervous system (CNS). GFAP levels in children with infantile autism were higher than those in normal children of the same age range. S-100 protein concentrations were similar in both groups. High levels of GFAP in combination with normal S-100 protein concentrations in CSF indicates reactive astrogliosis in the CNS. In conclusion, the sensitive ELISA described makes it possible to measure low levels of GFAP present in the CSF of children. Combined assays of GFAP and S-100 protein can be used to discriminate between acute and chronic brain disorders in children.