Rapid diagnosis of the systemic inflammatory response. syndrome by the transcription factor activation measuring method development concerning the inflammation

已知核因子-κB （NF-κB）转录因子家族在免疫系统调节中发挥重要作用。NF-κB被细菌和病毒抗原激活，导致促炎细胞因子和趋化因子的产生。通过全身性炎症反应综合征（SIRS）快速检测活化的NF-κB活化被认为对败血症患者的治疗至关重要。本研究的目的是评估两种方法的敏感性，电泳迁移量转移法（EMSA）和转录因子酶联免疫分析法（TF-ELISA）。TF-ELISA检测到重组人NF-κB p50 (rhp50) 25 ng和tnf α刺激的HeLa核蛋白5 mg， EMSA检测到约100 ng的rhp50和10 mg的HeLa核蛋白。tnf - elisa检测NF-κB的敏感性高于EMSA。但经判断，对于SIRS患者，TF-ELISA所需的3-6小时测量时间过长。因此，在医院实验室对活化NF-κB蛋白进行检测和定量，需要开发灵敏度更高、测量时间更短的新型分析方法。因此，我们开发了一种基于表面等离子体共振（SPR）的新型NF-κB分析仪，这是公认的最灵敏的直接光学检测方法之一。该方法可在15分钟内检测到NF-κB的纳摩尔浓度。目前，我们正在使用这种新的转录因子分析仪器对SIRS患者进行临床试验。

The nuclear factor-kappa B (NF-κB) family of transcription factors is known to play an important role in the regulation of the immune system. NF-κB is activated by bacterial and viral antigens, which leads to the production of proinflammatory cytokines and chemokines. The rapid detection of activated NF-κB activation by the systemic inflammatory response syndrome (SIRS) is considered to be crucial for the treatment of patients with septicemia. The aim of the present study was to evaluate the sensitivity of two methods, electrophoretic mobility shift assay (EMSA) and transcription factor enzyme-linked immunoassay (TF-ELISA). TF-ELISA detected 25 ng of recombinant human NF-κB p50 (rhp50) and 5 mg of TNFα-stimulated HeLa nuclear protein, while EMSA detected approximately 100 ng of rhp50 and 10 mg of HeLa nuclear protein. We found that the TF-ELISA was more sensitive than EMSA in detecting NF-κB. However, it was judged that the 3-6 hour measuring time required in TF-ELISA was excessively long for patients with SIRS. Therefore, the development of new analytical methods with improved sensitivity and, measurement time was necessary for the detection and quantification of activated NF-κB protein in the hospital laboratory. Consequently, we have developed a new NF-κB analyzer based on surface plasmon resonance (SPR), which is recognized as one of the most sensitive direct optical detection methods. This method can detect nanomolar concentrations of NF-κB within 15 minutes. At present, we are carrying out clinical trials using this new transcription factor analysis apparatus for SIRS patients.