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

核因子-kappaB(NF-κB)家族转录因子在免疫系统的调节中发挥重要作用。核因子-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。全身炎症反应综合征(SIRS)快速检测活化的NF-κB活性对败血症患者的治疗具有重要意义。本研究的目的是评价两种方法的敏感性，即凝胶迁移率改变分析(EMSA)和转录因子酶联免疫分析(TF-ELISA)。TF-ELISA检测到25 ng重组人核因子κB p50(RHp50)和5 mg肿瘤坏死因子α刺激的HeLa核蛋白，而EMSA检测到大约100 ng的rHp50和10 mg的HeLa核蛋白。我们发现，对于SIRS患者，TF-ELISA法检测NF-κB的灵敏度高于EMSA法，但对于SIRS患者，TF-ELISA法所需的3-6h检测时间过长。因此，开发新的分析方法，提高灵敏度和测量时间，对于在医院实验室检测和定量激活的NF-κB蛋白是必要的。因此，我们开发了一种新的基于表面等离子体共振的NF-κB分析仪，它被认为是最灵敏的直接光学检测方法之一。该方法可以在15分钟内检测到纳米分子浓度的核因子-κB。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.