Toxoplasma gondii Oocyst Protein
a technology of oocyst protein and toxoplasma gondii, which is applied in the field of toxoplasma gondii oocyst protein, can solve the problems of no test which can differentiate between oocyst ingestion and tissue cyst ingestion, and meat products containing tissue cysts which could infect consumers, and represent a potentially serious problem for the u.s. pork industry
Inactive Publication Date: 2009-08-27
HILL DOLORES E +3
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Benefits of technology
The patent describes the identification and use of specific proteins and peptides from the parasite Toxoplasma gondii that can be used for diagnosis and treatment of toxoplasmosis. These proteins can be used to detect antibodies in patients with the disease or to identify the parasite in biological samples. The invention also includes a method for identifying T. gondii oocysts in a subject and a diagnostic kit for the same. Overall, the invention provides new tools for the diagnosis and treatment of toxoplasmosis.
Problems solved by technology
Food animals, such as pigs, become infected by the same routes, resulting in meat products containing tissue cysts which could infect consumers.
Currently, there are no tests which can differentiate between oocyst ingestion versus tissue cyst ingestion as the infection route.
Increased consumer awareness of the potential risks of acquiring T. gondii from fresh pork products represents a potentially serious problem for the U.S. pork industry.
Food safety is a critical issue for the swine industry.
Method used
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[0057]Toxoplasma gondii RNA, DNA, and protein were extracted from unsporulated and sporulated oocysts, excysted sporozoites, tachyzoites, and bradyzoites stages using TRIZOL reagent (Gibco / BRL Life Technologies, Gaithersburg, Md.). For collection of parasite nucleic acids and proteins from unsporulated and sproulated oocysts, oocysyt walls were first disrupted by treating intact oocysts with 5.25% sodium hypochlorite in water for 30 minutes at room temperature. Oocysts were washed 3-4 times by centrifugation in water to remove the sodium hypochlorite, and were then disrupted by vortexing with 500 micron glass beads for 5 minutes. For collection of parasite nucleic acids and proteins from excysted sporozoites, tachyzoites, and bradyzoites, parasites were pelleted by centrifugation and lysed in 1-2 mls of TRIZOL reagent by repetitive pipetting (Hill, D. E., et al., “Specific detection of Neospora caninum oocysts in fecal samples from experimentally infected dogs using the polymerase c...
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Abstract
Recombinant proteins have been developed for the detection of Toxoplasma gondii oocyst proteins for example in biological fluids. Isolated DNA sequences which encode these proteins have also been developed. The DNA sequences may be inserted into recombinant DNA molecules such as cloning vectors or expression vectors for the transformation of cells and the production of the proteins.
Description
BACKGROUND OF THE INVENTION[0001]The present invention relates to an isolated 18.3 kDa protein, DGP5p+, specific for Toxoplasma gondii, which contains a transmembrane protein. The present invention also relates to DGP5p protein which lacks the transmembrane protein of DGP5p+; recombinant DGP5p (rDGP5p) protein; and the nucleic acid sequences which encode these proteins. The RNA which encodes DGP5p and rDGP5p can be used to specifically identify T. gondii oocysts through RT-PCR. The isolated and recombinant proteins can be used as reagents to detect antibodies in the serum of infected individuals.[0002]Toxoplasmosis, caused by the protozoan parasite Toxoplasma gondii, is one of the most common parasitic infections of man and other warm-blooded animals. It has been found worldwide from Alaska to Australia. Nearly one-third of humanity has been exposed to this parasite. In most adults it does not cause serious illness, but it can cause blindness and mental retardation in congenitally i...
Claims
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IPC IPC(8): G01N33/569C07K16/18
CPCA61K38/00A61K39/00C07K14/45G01N2469/00G01N33/56905G01N2333/45C07K16/20
Inventor HILL, DOLORES E.ZARLENGA, DANTE S.COSS, CATHLEENDUBEY, JITENDER P.
Owner HILL DOLORES E