Antigen cocktails, P35, and uses thereof

A composition comprising Toxoplasma gondii antigens P29, P35 and P66.

An isolated nucleic acid sequence represented by SEQ ID NO:

• 26.

A purified polypeptide having the amino acid sequence represented by SEQ ID NO:

• 27.

A method for detecting the presence of IgM antibodies to Toxoplasma gondii in a test sample comprising the steps of:

• a) contacting said test sample suspected of containing said IgM antibodies with a composition comprising P29, P35 and P66;
• and b) detecting the presence of said IgM antibodies.

A method for detecting the presence of IgM antibodies to Toxoplasma gondii in a test sample comprising the steps of:

• a) contacting said test sample suspected of containing said IgM antibodies with a composition comprising antigen P29, P35 and P66 for a time and under conditions sufficient for the formation of IgM antibody/antigen complexes;
• b) adding a conjugate to the resulting IgM antibody/antigen complexes for a time and under conditions sufficient to allow said conjugate to bind to the bound antibody, wherein said conjugate comprises an antibody attached to a signal generating compound capable of generating a detectable signal;
• and c) detecting the presence of IgM antibodies which may be present in said test sample by detecting a signal generated by said signal generating compound.

A method for detecting the presence of IgG antibodies to Toxoplasma gondii in a test sample comprising the steps of:

• a) contacting said test sample suspected of containing said IgG antibodies with a composition comprising P29, P30 and P35;
• and b) detecting the presence of said IgG antibodies.

A method for detecting the presence of IgG antibodies to Toxoplasma gondii in a test sample comprising the steps of:

• a) contacting said test sample suspected of containing said IgG antibodies with a composition comprising antigen P29, P30 and P35 for a time and under conditions sufficient for formation of IgG antibody/antigen complexes;
• b) adding a conjugate to resulting IgG antibody/antigen complexes for a time and under conditions sufficient to allow said conjugate to bind to bound antibody, wherein said conjugate comprises an antibody attached to a signal generating compound capable of generating a detectable signal;
• and c) detecting IgG antibodies which may be present in said test sample by detecting a signal generated by said signal generating compound.

A method for detecting the presence of IgM antibodies to Toxoplasma gondii in a test sample comprising the steps of:

• a) contacting said test sample suspected of containing said IgM antibodies with anti-antibody specific for said IgM antibodies for a time and under conditions sufficient to allow for formation of anti-antibody/IgM antibody complexes;
• b) adding a conjugate to resulting anti-antibody/IgM antibody complexes for a time and under conditions sufficient to allow said conjugate to bind to bound antibody, wherein said conjugate comprises a composition comprising P29, P35 and P66, each attached to a signal generating compound capable of generating a detectable signal;
• and c) detecting IgM antibodies which may be present in said test sample by detecting a signal generated by said signal generating compound.

A method for detecting the presence of IgG antibodies to Toxoplasma gondii in a test sample comprising the steps of:

• a) contacting said test sample suspected of containing said IgG antibodies with anti-antibody specific for said IgG antibodies for a time and under conditions sufficient to allow for formation of anti-antibody/IgG antibody complexes;
• b) adding a conjugate to resulting anti-antibody/IgG antibody complexes for a time and under conditions sufficient to allow said conjugate to bind to bound antibody, wherein said conjugate comprises a composition comprising P29, P30 and P35, each attached to a signal generating compound capable of generating a detectable signal;
• and c) detecting IgG antibodies which may be present in said test sample by detecting a signal generated by said signal generating compound.

A vaccine comprising:

• 1) Toxoplasma gondii antigens P29, P30 and P35 and 2) a pharmaceutically acceptable adjuvant.

A vaccine comprising:

• 1) Toxoplasma gondii antigens P29, P35 and P66 and 2) a pharmaceutically acceptable adjuvant.

A kit for determining the presence of IgM antibodies to Toxoplasma gondii in a test sample comprising:

• a) a composition comprising Toxoplasma gondii antigens P29, P35 and P66;
• and
• b) a conjugate comprising an antibody attached to a signal generating compound capable of generating a detectable signal.

A kit for determining the presence of IgG antibodies to Toxoplasma gondii in a test sample comprising:

• a) a composition comprising Toxoplasma gondii antigens P29, P30 and P35;
• and
• b) a conjugate comprising an antibody attached to a signal generating compound capable of generating a detectable signal.

A kit for determining the presence of IgM antibodies to Toxoplasma gondii in a test sample comprising:

• a) an anti-antibody specific for IgM antibody;
• and
• b) a composition comprising Toxoplasma gondii antigens P29, P35 and P66.

A kit for determining the presence of IgM antibodies to Toxoplasma gondii in a test sample comprising:

• a) an anti-antibody specific for IgM antibody;
• b) a conjugate comprising:
• 1) Toxoplasma gondii antigens P29, P35 and P66, each attached to 2) a signal generating compound capable of generating a detectable signal.

A kit for determining the presence of IgG antibodies to Toxoplasma gondii in a test sample comprising:

• a) an anti-antibody specific for IgG antibody;
• and
• b) a composition comprising Toxoplasma gondii antigens P29, P35 and P66.

A kit for determining the presence of IgG antibodies to Toxoplasma gondii in a test sample comprising:

• a) an anti-antibody specific for IgG antibody;
• b) a conjugate comprising:
• 1) Toxoplasma gondii antigens P29, P35 and P66, each attached to 2) a signal generating compound capable of generating a detectable signal.

A method for detecting the presence of IgM and IgG antibodies to Toxoplasma gondii in a test sample comprising the steps of:

• a) contacting said test sample suspected of containing said IgM and IgM antibodies with a composition comprising antigen P29, P30, P35 and P66 for a time and under conditions sufficient for the formation of IgM antibody/antigen complexes;
• b) adding a conjugate to the resulting IgM antibody/antigen complexes and IgG antibody/antigen complexes for a time and under conditions sufficient to allow said conjugate to bind to the bound IgM and IgG antibody, wherein said conjugate comprises an antibody attached to a signal generating compound capable of generating a detectable signal;
• and c) detecting the presence of IgM and IgM antibodies which may be present in said test sample by detecting a signal generated by said signal generating compound.

A method for detecting the presence of IgM and IgG antibodies to Toxoplasma gondii in a test sample comprising the steps of:

• a) contacting said test sample suspected of containing said IgM and IgG antibodies with anti-antibody specific for said IgM antibodies and said IgG antibodies for a time and under conditions sufficient to allow for formation of anti-antibody/IgM antibody complexes and anti-antibody/IgG antibody complexes;
• b) adding a conjugate to resulting anti-antibody/IgM antibody complexes and resulting anti-antibody/IgG antibody complexes for a time and under conditions sufficient to allow said conjugate to bind to bound antibody, wherein said conjugate comprises P29, P30, P35 and P66, each attached to a signal generating compound capable of generating a detectable signal;
• and c) detecting IgM and IgG antibodies which may be present in said test sample by detecting a signal generated by said signal generating compound.

The method of claim 34 wherein said antigen is derived from an organism selected from the group consisting of Borrelia burgdorferi, Schistosoma treponema, Toxoplasma gondii, Plasmodium vivax and Plasmodium falciparum.

A composition comprising the isolated nucleic acid sequence represented by FIG. 11 or a fragment thereof.

A composition comprising amino acids 1-135 of P35.

A method for distinguishing between acute and chronic Toxoplasmosis in a patient suspected of having either said acute or chronic Toxoplasmosis comprising the steps of:

• a) contacting a test sample, from said patient, with a composition comprising amino acids 1-135 of P35;
• and b) detecting the presence of IgG antibodies, presence of said IgG antibodies indicating acute Toxoplasmosis in said patient and lack of said IgG antibodies indicating chronic Toxoplasmosis in said patient.

A kit for distinguishing between acute and chronic Toxoplasmosis in a patient suspected of having either said acute Toxoplasmosis or said chronic Toxoplasmosis comprising:

• a) a composition comprising amino acids 1-135 of Toxoplasma gondii antigen P35;
• and
• b) a conjugate comprising an antibody attached to a signal generating compound capable of generating a detectable signal.

A kit for distinguishing between acute and chronic Toxoplasmosis in a patient suspected of having either said acute Toxoplasmosis or said chronic Toxoplasmosis comprising:

• a) an anti-antibody specific for IgG antibody;
• and
• b) a conjugate comprising amino acids 1-135 of Toxoplasma gondii antigen P35 attached to a signal generating compound capable of generating a detectable signal.