Fluorescent Nanoparticle-Based Indirect Immunofluorescence Microscopy for Detection ofMycobacterium tuberculosis

[摘要] A method of fluorescent nanoparticle-based indirect immunofluorescence microscopy(FNP-IIFM) was developed for the rapid detection ofMycobacterium tuberculosis. An anti-Mycobacterium tuberculosisantibody was used as primary antibody to recognizeMycobacterium tuberculosis, and then an antibody binding protein (Protein A) labeled withTris(2,2-bipyridyl)dichlororuthenium(II) hexahydrate (RuBpy)-doped silica nanoparticles wasused to generate fluorescent signal for microscopic examination. Prior to the detection, Protein Awas immobilized on RuBpy-doped silica nanoparticles with a coverage of∼5.1×102molecules/nanoparticle. With this method,Mycobacterium tuberculosisin bacterial mixture aswell as in spiked sputum was detected. The use of the fluorescent nanoparticles reveals amplifiedsignal intensity and higher photostability than the direct use of conventional fluorescent dye aslabel. Our preliminary studies have demonstrated the potential application of the FNP-IIFMmethod for rapid detection ofMycobacterium tuberculosisin clinical samples.

[学科分类] 基础医学