Development of a sensitive enzyme-linked immunosorbent assay for measurement of DNase I in human serum.
Development of a sensitive enzyme-linked immunosorbent assay for measurement of DNase I in human serum.
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DOI：
10.1016/j.cca.2009.03.005
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发表时间：
2009-05
期刊：
影响因子：
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通讯作者：
T. Nakajima;Rie Takagi;Yutaka Tajima;C. Makita;Y. Kominato;J. Kuribara;S. Ohshima;H. Tada;H. Tsurugaya;Yasuyuki Kobayashi;H. Takeshita;Yasuyuki Kawai;T. Yasuda
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文献类型：
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作者：
T. Nakajima;Rie Takagi;Yutaka Tajima;C. Makita;Y. Kominato;J. Kuribara;S. Ohshima;H. Tada;H. Tsurugaya;Yasuyuki Kobayashi;H. Takeshita;Yasuyuki Kawai;T. Yasuda
研究背景急性心肌梗塞（AMI）发病早期血清脱氧核糖核酸酶I（DNase I）活性升高。到目前为止，DNA酶I的活性已经通过单径向酶扩散（SRED）方法定量，不幸的是，该方法需要较长的孵育时间。因此，有必要开发另一种适合于在临床setting.METHODSA测定血清DNase I蛋白的浓度夹心ELISA建立了用于测量DNase I蛋白使用针对DNase I蛋白的多克隆抗体和生物素化的单克隆抗体用于随后的检测。本文测定了正常人和急性心肌梗塞患者血清DNase Ⅰ蛋白的含量，结果表明，本法与SRED法一样精确，但所需时间短。DNA酶I浓度与酶活性呈显著正相关（r=0.839; P<0.001）。AMI患者胸痛0-12 h内血清DNase I平均值明显高于健康人（P<0.001），并随时间延长而降低。此方法可能是一个有用的替代SRED作为辅助诊断AMI的基础上血清DNA酶I水平。
BACKGROUNDSerum deoxyribonuclease I (DNase I) activity was reported to increase in the early phase after onset of acute myocardial infarction (AMI). Up to now, DNase I activity has been quantified by the single radial enzyme diffusion (SRED) method, which unfortunately requires a long incubation time. Therefore it is necessary to develop another assay suitable for measurement of serum DNase I concentrations in a clinical setting.METHODSA sandwich ELISA was established for measurement of DNase I protein using a polyclonal antibody directed against DNase I protein and a biotinylated monoclonal for subsequent detection. Concentrations of serum DNase I protein were measured in healthy individuals and patients with AMI.RESULTSThis method was as precise as SRED, and took less time than SRED. A significant correlation was observed between DNase I concentration and enzyme activity (r=0.839; P<0.001). The average of serum DNase I in AMI patients within 0–12 h of chest pain was significantly higher than that in healthy individuals (P<0.001), and decreased with time.CONCLUSIONSWe have developed a sensitive ELISA capable of measuring DNase I protein concentrations. This method may be a useful alternative to SRED as an aid to diagnosis of AMI based on the serum DNase I level.