A Technique to Detect Extrapulmonary Tuberculosis via Superparamagnetic Nanoprobes
Published: January 31, 2024
Abstract
Source: Lee, C., et al. Synthesis, Characterization, and Application of Superparamagnetic Iron Oxide Nanoprobes for Extrapulmonary Tuberculosis Detection. J. Vis. Exp. (2020).
This video demonstrates a technique for the detection of extrapulmonary tuberculosis. Superparamagnetic iron oxide (SPIO) nanoprobes, conjugated to Mycobacterium-specific antibodies, are injected into a mouse previously infected with Mycobacterium bovis BCG, labeling the granuloma and providing a better imaging contrast for its detection.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. SPIO-MtbsAb synthesis Synthesize SPIO-conjugated 2,2'-(ethylenedioxy)bisethylamine (EDBE) using previously reported methods. Synthesize SPIO-EDBE-succinic anhydride (SA). Stir an alkaline solution (5 M NaOH; 10 mL)) of SPIO-EDBE and SA (1 g; 10 µmol) at room temperature for 24 h. <l…
Declarações
The authors have nothing to disclose.
Materials
| (benzotriazol-1-yloxy) tripyrrolidinophosphonium hexafluorophosphate | Sigma-Aldrich | ||
| 1-hydroxybenzotriazole | Sigma-Aldrich | ||
| epichlorohydrin, 2,2'-(ethylenedioxy)bis(ethylamine) | Sigma-Aldrich | ||
| Human monocytic THP-1 | |||
| M. bovis BCG | Pasteur Mérieux | Connaught strain; ImmuCyst Aventis | |
| MRI | GE medical Systems | 3.0-T, Signa | |
| NMR relaxometer | Bruker | NMS-120 Minispec | |
| SPECTRUM molecular porous membrane tubing, 12,000 -14,000 MW cut off | Spectrum Laboratories Inc | ||
| TB surface antibody- Polyclonal Antibody to Mtb | Acris Antibodies GmbH | BP2027 | |
| transmission electron microscope | JEOL | JEM-2000 EX II |
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Citar este artigo
A Technique to Detect Extrapulmonary Tuberculosis via Superparamagnetic Nanoprobes. J. Vis. Exp. (Pending Publication), e21895, doi: (2024).