A specific and sensitive ELISA for measuring S-100b in cerebrospinal fluid
A specific and sensitive ELISA for measuring S-100b in cerebrospinal fluid
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DOI：
10.1016/s0022-1759(97)00050-1
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发表时间：
1997-06-23
影响因子：
2.2
通讯作者：
Thompson, EJ
中科院分区：
文献类型：
--
作者：
Green, AJE;Keir, G;Thompson, EJ
建立了一种灵敏、简便、特异的S-100b夹心ELISA试剂盒。该方法包括将单克隆抗s -100b抗体结合到微滴板壁上。该捕获抗体随后与S-100b标准、对照或脑脊液（CSF）形式的患者样本孵育。孵育后，清洗微滴板，加入辣根过氧化物酶标记的多克隆抗s -100b（检测抗体）。检测抗体结合到微滴板上的量与样品中S-100b的量成正比。该方法检测下限为0.04 ng/ml，与密切相关的多肽S-100a的反应性< 0.006%。在S-100b浓度分别为0.38和0.8 ng/ml时，该方法的批内平均精密度分别为9.3%和5.6%。S-100b浓度分别为0.12和0.34 ng/ml时，批间精密度分别为8.9和8.1%。添加0.5 ng/ml时，CSF中S-100b的回收率为94%，CV为8.5%。使用预涂微滴板，该测定可在不到5小时内完成，因此可在临床实验室中常规使用。使用该ELISA分析了154份脑脊液样本，发现19%的样本水平升高。脑出血或中枢神经系统恶性肿瘤患者的血药浓度最高。无神经疾病证据的个体S-100b浓度低于0.4 ng/ml。只有5%的多发性硬化症患者发现CSF S-100b浓度升高。从一名受感染的住院分流患者身上采集的一系列脑脊液样本显示出急剧下降，表明S-100b可以迅速清除。(C) 1997爱思唯尔科学有限公司
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.