Klebsiella pneumoniae is a common human pathogen that causes several nosocomial chronic infections. Many methods have been reported to detect Klebsiella pneumonia; however, a simple, rapid, and visual method remains to be developed. Here, we developed a rapid, visual detection method employing a nanoparticle immunosensor (NPIS) with a sensitivity of 8 CFU mL^-1. Two monoclonal antibodies (mAbs) recognizing different antigenic determinants on the surface of Klebsiella pneumonia were screened by an indirect enzyme-linked immunosorbent assay (ELISA), Western blotting, and mass spectrometry assay. The mAb 1E6 was conjugated to magnetic nanoparticles (MNPs) to generate the immuno-magnetic nanoparticles (IMNPs), whereas the horseradish peroxidase (HRP)-mAb probe was synthesized by conjugating the mAb 2F1 with HRP. Thereafter, the NPIS was developed based on the IMNPs and the HRP-mAb probe, and the total assay time for the detection of Klebsiella pneumoniae was ∼60 min. A rapid, sensitive method was developed for screening Klebsiella pneumoniae in clinical samples. The NPIS developed in this study can detect Klebsiella pneumoniae without the enrichment of bacteria and the extraction of the genome; therefore, it can be used as a primary screening method to compliment other methods in the detection of Klebsiella pneumoniae.

中文翻译：

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使用磁性纳米颗粒和基于辣根过氧化物酶探针的免疫传感器快速，直观地检测肺炎克雷伯菌。

肺炎克雷伯菌是一种常见的人类病原体，可引起多种医院内慢性感染。已有许多方法检测克雷伯菌肺炎; 但是，仍然需要开发一种简单，快速和可视化的方法。在这里，我们开发了一种快速，直观的检测方法，该方法采用了灵敏度为8 CFU mL ^-1的纳米粒子免疫传感器（NPIS）。两种识别肺炎克雷伯菌表面抗原决定簇的单克隆抗体（mAb）通过间接酶联免疫吸附测定（ELISA），Western印迹和质谱测定法进行筛选。将mAb 1E6与磁性纳米颗粒（MNP）偶联以生成免疫磁性纳米颗粒（IMNP），而辣根过氧化物酶（HRP）-mAb探针是通过将mAb 2F1与HRP偶联而合成的。此后，基于IMNP和HRP-mAb探针开发了NPIS，检测肺炎克雷伯菌的总检测时间约为60分钟。开发了一种快速，灵敏的方法来筛查肺炎克雷伯菌在临床样品中。这项研究开发的NPIS可以检测肺炎克雷伯菌，而无需富集细菌和提取基因组。因此，它可以作为对肺炎克雷伯菌检测中其他方法的补充的主要筛选方法。