Rapid, Visual Detection of Klebsiella pneumoniae Using Magnetic Nanoparticles and an Horseradish Peroxidase-Probe Based Immunosensor

摘要

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 similar to 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.