Diagnosis of clinical toxoplasmosis remains a challenge, thus limiting the availability of human clinical samples. Though murine models are an approximation of human response, their definitive infection status and tissue availability make them critical to the diagnostic development process. Hydrogel mesh nanoparticles were used to concentrate antigen to detectable levels for mass spectrometry. Seven Toxoplasma gondii isolates were used to develop a panel of potential peptide sequences for detection by parallel reaction monitoring (PRM) mass spectrometry. Nanoparticles were incubated with decreasing concentrations of tachyzoite lysate to explore the limits of detection of PRM. Mice whose toxoplasmosis infection status was confirmed by quantitative real-time PCR had urine tested by PRM after hydrogel mesh concentration for known T. gondii peptides. Peptides from GRA1, GRA12, ROP4, ROP5, SAG1, and SAG2A proteins were detected by PRM after nanoparticle concentration of urine, confirming detection of T. gondii antigen in the urine of an infected mouse.
|Number of pages||9|
|Journal||Nanomedicine: Nanotechnology, Biology, and Medicine|
|State||Published - Feb 2018|
Bibliographical noteFunding Information:
Funding from INNOVATE PERU (137-PNICP-PIAP) allowed for the collaboration of Universidad Peruana Cayetano Heredia and Universidad Nacional Mayor de San Marcos as well as equipment, training and materials. Dr. Gilman's 1D43TW010074-01 supported the training of many of the Peruvian authors. This study was partially funded by the following grants to Dr. Liotta: National Institutes of Health NCI R33CA173359 and R33CA206937, NIAID R21AI099851 and R21AI117425, and NIAMS 1R01AR068436. Drs. Liotta and Luchini are inventors on patents related to the nanoparticles. Ceres Nanosciences licensed the rights of these patents that are owned by George Mason University. Drs. Liotta and Luchini own shares of Ceres Nanosciences.
- Multiple reactions monitoring
- Toxoplasma gondii