Journal of Biomedicine and Biotechnology 
Volume 2007 (2007), Article ID 89364, 9 pages
doi:10.1155/2007/89364
Research Article

Fluorescent Nanoparticle-Based Indirect Immunofluorescence Microscopy for Detection of Mycobacterium tuberculosis

Dilan Qin,1,2 Xiaoxiao He,1,2 Kemin Wang,1,2 Xiaojun Julia Zhao,3 Weihong Tan,1,2 and Jiyun Chen1,2

 1State Key Laboratory of Chemo/Biosensing and Chemometrics, Biomedical Engineering Center, College of Chemistry and Chemical Engineering, Hunan University, Changsha, Hunan 410082, China
2Key Laboratory for Bio-Nanotechnology and Molecule Engineering of Hunan Province, Hunan University, Changsha 410082, China
3Department of Chemistry, University of North Dakota, Grand Forks 58202, ND, USA
Received 24 March 2007; Revised 10 July 2007; Accepted 10 October 2007

Recommended by Marek Osinski

Abstract

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