Institute of Biomedical Engineering, College of Life Science, Qingdao University, Qingdao, 266071, China.
Institute of Biomedical Engineering, College of Life Science, Qingdao University, Qingdao, 266071, China.
Anal Chim Acta. 2024 Feb 22;1291:342225. doi: 10.1016/j.aca.2024.342225. Epub 2024 Jan 9.
Endotoxin detection is important for determining bacterial contamination and infection in fields of food, pharmaceutical and clinical disease diagnosis. The horseshoe crab deformed cell lysate analysis is regarded as the gold-standard method, but the endangered and high-cost horseshoe crab animals required in sensing process further raise animal ethical issues and hinder their applications. The colorimetric methods based on nanozymes are simple and economical, but the low selectivity and sensitivity are still the bottleneck for their further application. Herein, we successfully developed a phenylboronic acid functionalized iron-based nanozyme with higher selectivity and highly catalytic activity for endotoxin sensing. The as-prepared colorimetric sensor using the obtained nanozyme as sensing probes shows a good linear relationship for endotoxin sensing in the range of 1-20 μg mL, with a LOD = 0.42 μg mL, along with good selectivity and reproducibility. The sensor can also be well applied to detecting endotoxin in practical samples such as beer and serum. Moreover, the parameters including time and temperature which could affect the endotoxin release from E. coli were also studied and optimized, based on the relationship between endotoxin and Gram-negative bacteria, the as-prepared sensor achieves the qualitative and quantification of E. coli.
内毒素检测对于确定食品、制药和临床疾病诊断领域的细菌污染和感染非常重要。马蹄蟹变形细胞裂解物分析被认为是金标准方法,但在传感过程中需要濒危且昂贵的马蹄蟹动物,这进一步引发了动物伦理问题,并阻碍了它们的应用。基于纳米酶的比色法简单且经济,但选择性和灵敏度低仍然是其进一步应用的瓶颈。在此,我们成功开发了一种基于苯硼酸的铁基纳米酶,具有更高的选择性和高度的催化活性,可用于内毒素传感。使用所获得的纳米酶作为传感探针制备的比色传感器在 1-20μg mL 的范围内对内毒素传感表现出良好的线性关系,LOD=0.42μg mL,具有良好的选择性和重现性。该传感器还可以很好地应用于检测实际样品如啤酒和血清中的内毒素。此外,还研究和优化了可能影响大肠杆菌中内毒素释放的时间和温度等参数,基于内毒素与革兰氏阴性菌的关系,制备的传感器实现了对大肠杆菌的定性和定量检测。