A specific and sensitive ELISA for measuring S-100b in cerebrospinal fluid

被引:62
作者
Green, AJE
Keir, G
Thompson, EJ
机构
[1] Department of Neuroimmunology, National Hospital for Neurology and Neurosurgery, London WC1N 3BG, Queen Square
关键词
S-100b; ELISA; cerebrospinal fluid; central nervous system;
D O I
10.1016/S0022-1759(97)00050-1
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A sensitive, simple and specific sandwich ELISA for S-100b is described. This method involves the binding of a monoclonal anti-S-100b antibody to the wall of a microtitre plate. This capture antibody is subsequently incubated with S-100b standard, control or patient sample in the form of cerebrospinal fluid (CSF). After incubation, the microtitre plate is washed and horseradish peroxidase-labelled polyclonal anti-S-100b is added (detector antibody). The amount of detector antibody bound to the microtitre plate is proportional to the amount of S-100b in the sample. The assay has a lower limit of detection of 0.04 ng/ml and shows < 0.006% reactivity with the closely related polypeptide S-100a. The assay has a mean within-batch precision of 9.3 and 5.6% at S-100b concentrations of 0.38 and 0.8 ng/ml, respectively. The between batch precision is 8.9 and 8.1% at S-100b concentrations of 0.12 and 0.34 ng/ml, respectively. The recovery of S-100b from CSF spiked with 0.5 ng/ml was 94% with a CV of 8.5%. The assay may be completed in less than 5 h using precoated microtitre plates, thus lending itself to routine use in clinical laboratories. Using this ELISA, 154 CSF samples were analysed and 19% of samples were found to have elevated levels. The highest levels were found in patients with cerebral haemorrhage or central nervous system malignancy. S-100b concentrations from individuals without evidence of neurological disease were found to be less than 0.4 ng/ml. Only 5% of patients with multiple sclerosis were found to have elevated CSF S-100b concentrations. Serial CSF samples taken from a patient with an infected in-dwelling shunt showed a dramatic decline, suggesting that S-100b is rapidly cleared. (C) 1997 Elsevier Science B.V.
引用
收藏
页码:35 / 41
页数:7
相关论文
共 16 条
[1]   THE S-100 PROTEIN IN CEREBROSPINAL-FLUID - A SIMPLE ELISA METHOD [J].
AURELL, A ;
ROSENGREN, LE ;
WIKKELSO, C ;
NORDBERG, G ;
HAGLID, KG .
JOURNAL OF THE NEUROLOGICAL SCIENCES, 1989, 89 (2-3) :157-164
[2]   THE S-100 - A PROTEIN FAMILY IN SEARCH OF A FUNCTION [J].
FANO, G ;
BIOCCA, S ;
FULLE, S ;
MARIGGIO, MA ;
BELIA, S ;
CALISSANO, P .
PROGRESS IN NEUROBIOLOGY, 1995, 46 (01) :71-82
[3]  
GRIFFIN WST, 1995, J NEUROCHEM, V65, P228
[4]  
GRIFFIN WST, 1993, METH NEUROSCI, V17, P267
[5]  
KATO K, 1985, JPN J CANCER RES, V76, P856
[6]   S100AO (ALPHA-ALPHA) PROTEIN IS MAINLY LOCATED IN THE HEART AND STRIATED MUSCLES [J].
KATO, K ;
KIMURA, S .
BIOCHIMICA ET BIOPHYSICA ACTA, 1985, 842 (2-3) :146-150
[7]   SENSITIVE ENZYME-IMMUNOASSAY FOR S-100 PROTEIN - DETERMINATION IN HUMAN CEREBROSPINAL-FLUID [J].
KATO, K ;
NAKAJIMA, T ;
ISHIGURO, Y ;
TENHOSHIMARU, M .
BIOMEDICAL RESEARCH-TOKYO, 1982, 3 (01) :24-28
[8]   MYELIN BASIC-PROTEIN AND S-100 ANTIGEN IN CEREBROSPINAL-FLUID OF PATIENTS WITH MULTIPLE-SCLEROSIS IN THE ACUTE PHASE [J].
MASSARO, AR ;
MICHETTI, F ;
LAUDISIO, A ;
BERGONZI, P .
ITALIAN JOURNAL OF NEUROLOGICAL SCIENCES, 1985, 6 (01) :53-56
[9]   S-100 ANTIGEN IN CEREBROSPINAL-FLUID AS A POSSIBLE INDEX OF CELL INJURY IN THE NERVOUS-SYSTEM [J].
MICHETTI, F ;
MASSARO, A ;
RUSSO, G ;
RIGON, G .
JOURNAL OF THE NEUROLOGICAL SCIENCES, 1980, 44 (2-3) :259-263
[10]   NERVOUS SYSTEM-SPECIFIC S-100 ANTIGEN IN CEREBROSPINAL-FLUID OF MULTIPLE-SCLEROSIS PATIENTS [J].
MICHETTI, F ;
MASSARO, A ;
MURAZIO, M .
NEUROSCIENCE LETTERS, 1979, 11 (02) :171-175