WO2012005563A1

WO2012005563A1 - Uses of biological compounds from toxoplasma sp and related compounds

Info

Publication number: WO2012005563A1
Application number: PCT/MY2010/000141
Authority: WO
Inventors: Ramaha Noordin; Geita Saadaatnia; Zeehaida Mohamed
Original assignee: Universiti Sains Malaysia
Priority date: 2010-07-07
Filing date: 2010-08-11
Publication date: 2012-01-12
Also published as: MY197339A

Abstract

The present invention relates to a plurality of biological compounds, derived from Toxoplasma sp. There is provided a method for detecting substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection. In particular the present invention uses excretory- secretory proteins produced by Toxoplasma sp. as a biomarker of infection. The present invention also provides for the use of these biological compounds in a medicament for management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom, as well as their use in the manufacture of a product for diagnosis, management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom.

Description

USES OF B!OLOGfCAL COMPOUNDS FROM TOXOPLASMA SP AND

RELATED COMPOUNDS

Background of the Invention Field of the Invention

This invention relates to uses of biological compounds, and more particularly to •the uses of biological compounds from Toxoplasma sp and other related compounds.

Description of Related Arts

Toxoplasma gondii is an obligate intracellular protozoan parasite. Infection with this parasite is widespread and important in humans, especially pregnant women and immunosuppressed patients. During pregnancy, primary infection of women is associated with fetal infection, which can lead to abortion, neonatal malformations and a multitude of other manifestations (Williams et at., 1981 ). The main objective of all diagnostic efforts in pregnant women is to determine whether the infection is acute (active) or chronic (Lecordier et al., 2000). In immunocompromised individuals (e.g. those with AIDS, neoplastic diseases, bone marrow or heart transplant recipients), toxoplasmosis (acute or, most often, reactivation of chronic infection) may be life-threatening (Frenkel, 1988). Unless treated, this can result in severe morbidity and often death. These severe sequelae underline the importance of appropriate laboratory investigations in^■ potential cases of acute/reactivated Toxoplasma infection. Seronegative patients are susceptible to infection and should be given advice on precautions to avoid exposure to Toxoplasma (Porter et al., 1992).

Diagnosis of acute toxoplasmosis still poses a lot of challenges, and a panel of tests is required for confirmation of toxoplasmosis. In most laboratories, the routine diagnosis of toxoplasmosis is based on serological methods, using many different kinds of available commercialized ELISA kits. Most of these kits employ ^' soluble antigen .from the parasite and thus vary in specificities and sensitivities, . which poses challenges in interpretation of tests results.

It has been observed that excretory-secretory proteins are highly immunogenic during both human and experimental infections (Darcy et al-, 1988; Decoster et al., 1988). They are therefore good candidates for investigation into new infection markers. The present invention provides Toxoplasma gondii compounds as new antigen candidates for development of specific and sensitive immunoassays and vaccine for toxoplasmosis and Toxoplasma infection in general.

References

• Darcy, F., Deslee, D., Santoro, F., Charif, H., Auriault, C, Decoster, A., Duquesne, V., & Capron, A. (1988). Induction of a protective antibody-dependent response against toxoplasmosis by in vitro excreted/secreted antigens from tachyzoites of Toxoplasma gondii. Parasite Immunol, 10{5), 553-567.

• Decoster, A., Darcy, F., & Capron, A. (1988). Recognition of Toxoplasma gondii excreted and secreted antigens by human sera from acquired and congenital toxoplasmosis: identification of markers of acute and chronic infection. Clin Exp Immunol, 73(3), 376-382.

• Frenkel, J. K. (1988). Pathophysiology of toxoplasmosis. Parasitoi Today, 4(10), 273-278.

• Lecordier, L, Fourmaux, M. P., Mercier, C., Dehecq, E., Masy, E., & Cesbron-Delauw, M. F. (2000). Enzyme-linked immunosorbent assays using the recombinant dense granule antigens GRA6 and GRAT of Toxoplasma gondii for detection of immunoglobulin G antibodies. Clin Diagn Lab Immunol, 7(4), 607-61 1.

• Porter, S. B., & Sande, M. A. ( 992). Toxoplasmosis of the central nervous system in the acquired immunodeficiency syndrome. N Engl J Med, 327(23), 1643-1648.

• Williams, K. A.,^' Scott, J. M., Macfarlane, D. E., Williamson, J. M., Elias-Jones, T. F., & Williams, H. (1981 ). Congenital toxoplasmosis: a prospective survey in the West of Scotland. J Infect, 3(3), 219-229. Summary of Invention

It is an object of the present invention to provide compounds for the detection of substances indicating infection by Toxoplasma sp, in a biological sample.

It is also an object of the present invention to provide a medicament for management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals.

. . .

It is another object of the present invention to provide a compound for use in the manufacture of a product for diagnosis, management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals. Accordingly, these objectives may be achieved by following the teachings of the present invention. There is provided a method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection, comprising the steps of: firstly, contacting a biomarker with the biological sample under conditions and over a time sufficient to form a complex with biological products, in the biological sample; then, detecting the presence or absence of said complex in the biological sample; and then optionally, analysing the result of said detection by comparing the result with a comparison sample or a biological reference sample or both; wherein said biomarker is characterized by, derived from or made using an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID • NO. 14, or any combination thereof; or an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO. 6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 10; antibodies to SEQ ID NO. 12, antibodies to SEQ.ID NO. 14, antibodies to SEQ ID NO. 1 , antibodies to . SEQ. ID NO. 3, antibodies to SEQ ID NO. 5, antibodies to SEQ ID NO. 7 antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO. 11 , antibodies to SEQ ID NO. 13, or any combination thereof; or a nucleotide sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 1 , SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 11 , SEQ ID NO. 3, or any combination thereof; or any combination thereof. The present invention also provides for the use of these biological compounds in a medicament for management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals. The present invention also relates to the use of these biological compounds in the manufacture of a product for diagnosis, management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals._.

Brief Description of the Drawings

The features of the invention will be more readily understood and appreciated from the following detailed description when read in conjunction with the accompanying drawings of the preferred embodiment of the present invention, in which:

• Fig. 1 is a diagram showing the immunoblot patterns of excretory secretory antigens of in-vitro grown of T. gondii recognized by different serum samples when probed with peroxidase conjugated anti-human IgM antibody, wherein: a) low molecular weight marker, b) sample from Group I sera (probable active toxoplasmosis), c) sample from Group II sera (chronic toxoplasmosis), d) sample from Group III sera (normal serum), e) sample from Group IV sera (other infections) .

• Fig. 2 is a diagram showing the immunoblot patterns of excretory secretory antigens of in-vitro grown of T. gondii recognized by different serum samples when probed with peroxidase , conjugated anti-human IgA antibody, wherein a) low molecular weight marker, b) sample from Group I sera (probable active toxoplasmosis), c) sample from Group II sera (chronic toxoplasmosis), d) sample from Group III sera (normal serum), e) sample from Group IV sera (other infections)

• Fig. 3 is a diagram showing the immunoblot patterns of excretory secretory · antigens of in-vivo grown T. gondii recognized by different serum samples when probed with peroxidase conjugated anti-human IgM antibody, wherein a) low molecular weight marker, b) sample from Group I sera (probable active toxoplasmosis), c) sample from Group II sera (chronic toxoplasmosis), d) sample from Group III sera (normal serum), e) sample from Group IV sera (other infections)

• Fig. 4 is a diagram showing the immunoblot patterns of excretory secretory antigens of in-vivo grown T. gondii recognized by different serum samples when probed with peroxidase conjugated anti-human IgA antibody, wherein a) low molecular weight marker, b) sample from Group I sera (probable active toxoplasmosis), c) sample from Group II sera (chronic toxoplasmosis), d) sample from Group III sera (normal serum), e) sample from Group IV sera (other infections) · Sequence Listing (according to PCT Standard ST. 25) of the amino acid sequences of the 10, 12, 20 and 30 kDa proteins (SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14) and the nucleotide sequence of their DNA coding sequences (SEQ ID NO. 1 , SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 1 1 , SEQ ID NO. 13).

Detailed Description of the Invention

As required, detailed embodiments of the present invention are disclosed herein; however, it is to be understood that the disclosed embodiments^' are merely examples of the invention, which may be embodied in various forms. Therefore, specific structural and functional details disclosed herein'are not to be interpreted as limiting but merely as a basis for claims. It should be understood that the drawings and detailed description thereto are not intended to limit the invention to the particular form disclosed, but on the contrary, the invention is to cover all modification, equivalents and alternatives falling within the spirit and scope of the 5 present invention as defined by the appended claims. As used throughout this application, the word "may" is used in a permissive sense (i.e., meaning having the potential to), rather than the mandatory sense (i.e., meaning must). Similarly, the words "include," "including," and "includes" mean including, but not limited to. Further, the words "a" or "an" mean "at least one" and the word "plurality" means0 one or more, unless otherwise mentioned. Where the "abbreviations of technical terms are used, these indicate the commonly accepted meanings as known in the technical field. For ease of reference, common reference numerals will be used throughout the figures when referring to the same or similar features common to the figures. The present invention will now be described with reference to Figs. 1-45 and the accompanying Sequence Listing.

In the present invention, there is provided a method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp;0 infection, comprising the steps of: firstly, contacting a biomarker with the biological sample for a time and under conditions sufficient to form a complex with biological products in the biological sample; then, detecting the presence or absence of said complex in the biological sample; and then optionally, analysing the result of said detection by comparing the result with a comparison sample or a biological reference sample or both; wherein said biomarker is characterized by, derived from or made using: an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ^' ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof; or an amino, acid sequence substantially identical, in part or in whole, to a sequence^' selected from the group consisting of ^{' '} . antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO.

6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 10, antibodies to- SEQ ID NO. 12, antibodies to SEQ ID_. NO. 14, antibodies to SEQ ID NO. 1 , antibodies to SEQ ID NO. 3, antibodies to SEQ ID NO. 5, antibodies to SEQ ID NO. 7 antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO. 1 1 , antibodies to SEQ ID NO. 13, or any combination thereof; or a nucleotide sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 1 , SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 1 1 , SEQ ID NO. 13, or any combination thereof; or any combination thereof; and wherein said method may optionally be carried out using a detection kit. SEQ ID NO. 1 , SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 8, SEQ ID NO. 9, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 13, and SEQ ID NO. 14 are shown in the Sequence Listing.

More preferably, the Toxoplasma sp. is Toxoplasma gondii. While the method of the present invention may be used for various types of biological sample, it is preferred that ^'the biological sample is whole blood or blood serum or cerebrospinal fluid or eye fluid from a human or animal.

The method may be used to detect a variety of different substances produced by or present in Toxoplasma sp., but in one of the preferred embodiments, the substances produced by or present in Toxoplasma sp. are polypeptides. More preferably, the polypeptides comprise an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof.

In one of the embodiments of the present invention, the substances produced by or present in a human or animal body in response to Toxoplasma sp. infection, which- are detected by said method, are antibodies. More preferably, the antibodies are antl-Toxoplasma IgM or IgA or IgG or any combinations thereof.

It is possible that the biomarker may comprise a compound characterized by, derived from or made using an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting o SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof and wherein the biological products in the biological sample are antibodies produced in a human or animal body in response to Toxoplasma sp. infection.

. In one of the embodiments of the present invention, the biomarker ma comprise a compound characterized by, derived from or made Using an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO. 6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 10, antibodies to SEQ ID NO. 12, antibodies to SEQ ID NO. 14, antibodies to SEQ ID NO. 1 , antibodies to SEQ ID NO. 3, antibodies to SEQ ID NO. 5, antibodies to SEQ ID NO. 7, antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO. 1 1 , antibodies to SEQ ID NO.13, or any combination thereof and wherein the biological products in the biological sample are Toxoplasma sp. antigens.

The biomarker may comprise a compound characterized by, derived from or made using a nucleotide sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 1 , SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 1 1 , SEQ ID NO. 13, or any combination thereof and wherein the biological products in the biological sample are antibodies produced in a human or animal body in response to Toxoplasma sp. infection.

For said method, the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6; SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof may include native, or recombinant forms. Said method may also be carried out in an embodiment wherein the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof includes antigenic epitopes thereof. In one embodiment of said method, the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of the antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO. 6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 10, antibodies to SEQ ID NO. 12, antibodies to SEQ ID NO. 14, antibodies to SEQ ID NO. 1 , antibodies to SEQ ID NO. 3, antibodies to SEQ ID NO. 5, antibodies to .SEQ ID NO. 7, antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO. 11 , antibodies to SEQ ID NO.13, or any combination thereof includes any form such as monoclonal, recombinant or polyclonal. antibodies.

The method of the present invention may be carried out in a variety of circumstances including in a laboratory or optionally, using the detection kit. For example, the method of the present invention may be carried out using ELISA tests, dot blot, Western blot, flow-through rapid tests, lateral flow rapid tests, or biosensor tests. The detection kit for carrying out said method may comprise: the biomarker; a detection reagent for detecting said complex in the biological sample; optionally the comparison sample; and optionally the biological reference sample; wherein said biomarker, detection reagent, comparison sample and biological reference sample are present in amounts sufficient to perform said method

The comparison sample used in the method may comprise substances able to specifically bind to the biomarker, e.g. antibodies to the biomarker. Contacting the biomarker with said comparison sample may therefore provide a reference to be compared with the result obtained from said detection. Alternatively, the result is compared to the biological reference sample. Preferably, the biological reference sample is a sample that lacks substances able to bind to the biomarker. The step of comparing the result with a comparison sample or a biological reference sample or both is optional, and it will be obvious to persons skilled in the art that the method of the present.ihvention may be carried out even without said comparison.

The present invention also relates to a medicament for management or prophylaxis of .Toxoplasma sp. infection or a disease resulting therefrom in humans or animals, having a medicinal compound comprising: an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof; or an amino acid sequence substantially identical, in part or in wholes to a sequence selected from the group consisting of antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies^' to SEQ ID NO. 6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 10, antibodies to SEQ ID NO. 12, antibodies to SEQ ID NO. 14,· antibodies to SEQ ID NO. 1 , antibodies to SEQ ID NO. 3, antibodies to SEQ ID NO. 5, antibodies to SEQ ID NO. 7 antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO. 1 , antibodies to SEQ ID NO. 13, or any combination thereof; or a nucleotide sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 1 , SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 1 1 , SEQ ID NO. 13, or any combination thereof; or any combination thereof; wherein the medicinal compound may be present alone or in combination with other medicinal substances, and wherein said medicament may further include an acceptable carrier, diluent, or excipient.

In said medicament, the amino acid sequence substantially identical, in. part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof, may include native or recombinant forms. In one embodiment of the medicament, the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof, includes antigenic epitopes thereof. In one embodiment, the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of the antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO. 6, antibodies to SEQ ^'ID NO. 8, antibodies to SEQ ID NO, 10, antibodies to SEQ ID NO. 12, antibodies to SEQ ID NO. 14, antibodies to SEQ ID NO, 1 , antibodies to SEQ }D 1 1 . ^{· '} . - . ·^■ · NO: 3, antibodies to SEQ ID NO. 5, antibodies to SEQ ID NO. 7, antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO, 1 1 , antibodies to SEQ ID NO. 3, or any combination thereof, includes any form such as monoclonal, recombinant or polyclonal antibodies. Preferably, , the carrier, diluent, or excipient are. of pharmaceutical grade and suitable for use in the formulation of said medicament for administration to a human or animal. In the preferred embodiment, said disease is toxoplasmosis or complications thereof. The medicament may be, for example, a vaccine for prevention of said disease. The present invention also relates to a product, or a compound for use in .the manufacture of a product, for diagnosis, management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals, wherein the compound is derived from, made using or characterized by: an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof; or an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO. 6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 10, antibodies to SEQ ID NO. 12, antibodies to SEQ ID NO. 14, antibodies to SEQ ID NO. 1 , antibodies to SEQ ID NO. 3, antibodies to SEQ ID NO. 5, antibodies to SEQ ID NO. 7 antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO. 1 1 , antibodies to SEQ ID NO. 3, or any combination thereof; or a nucleotide sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 1 , SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO, 7, SEQ ID NO. 9, SEQ ID NO. 1 1 , SEQ ID NO. 13, or any combination thereof. Preferably, the disease is toxoplasmosis or complications thereof. Said compound may, for example, be used in the manufacture of a vaccine against toxoplasmosis. In one embodiment, said product includes gel electrophoresis bands, or electrophoresis bands transferred onto a membrane, or any combination thereof. For instance, said compound may be resolved by gel electrophoresis into bands and transferred by electro blotting onto a nitrocellulose membrane by western blot procedure, The membrane bearing the bands may be used, for example, in immunoblot assays for diagnosing toxoplasmosis.

The product, or compound for use in the manufacture of a product, for diagnosis, management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals, may be derived from, made using or characterized by the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof, including native or recombinant forms. In one embodiment, the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof, includes antigenic epitopes thereof. Said product or compound may be derived from, made using or characterized by the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of the antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO. 6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 10, antibodies to SEQ ID NO. 12, antibodies to SEQ ID NO. 14, antibodies to SEQ ID NO. 1 , antibodies to SEQ ID NO. 3, antibodies to SEQ ID NO. 5, antibodies to SEQ ID NO. 7 antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO. 1 , antibodies to SEQ ID NO. 13, or any combination thereof, including any form such as monoclonal, recombinant or polyclonal antibodies. Example

Below is an example of a study of the compounds of SEQ ID NO. 1 , SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 8, SEQ ID NO. 9 and SEQ ID NO. 10 from which the advantages of the present invention may be more readily understood. It is to be understood that the following example is for illustrative purpose only and should not be construed to limit the invention in any way. Materials

Firstly, excretory-secretory proteins (ESA) were prepared from in-vivo as well as in-vitro grown Toxoplasma gondii tachyzoites. In-vivo grown tachyzoites were grown in female Swiss albino mice (6 weeks old) infected with virulent RH strain of T. gondii. African green monkey kidney cells (VERO cell line) in DMEM culture medium were used for growing the in-vitro grown tachyzoites.

Blood serum samples used in this study were taken from three categories of individuals: Group I (individuals with probable active infection), anti-Toxop/asma lgM+, lgG+ or lgM+, IgG-, n = 75; Group II (individuals with chronic infection), anti- Toxoplasma IgM-, lgG+, n = 20; Group III (normal healthy individuals), anti-Toxoplasma IgM-, IgG-, n=20; Group IV (samples from individuals having other infections namely malaria, filariasis, leptospirosis, -dengue and amoebic liver abscess), anti-Toxop/asma IgM-, IgG-, n= 30.

SDS-PAGE and Western blotting

The ESA was dissolved in sample buffer (1 :1 ratio) containing 28.6% SDS and 4.76% 2- mercaptoethanol, boiled for 10 minutes and subsequently centrifuged for 3 minutes at 12000 g. It was then subjected to 12% sodium dodecyl sulfate polyacrylamid gel electrophoresis (SDS-PAGE) using Mini-Protein 3 cell equipment (Biorad, USA) in running buffer containing 0.3% tris, 1.44% glycin and 0.1 % SDS with a pH of 8.3. ESA with protein concentration of 20 pg per lane was evenly loaded, and electrophoresis was ran at room temperature with a constant current of 100 V for 90 minutes. Transfer buffer was prepared from a mixture of 0.3% tris, 1 .44% glycin and 20% methanol. The gel was then transferred to a nitrocellulose paper (NCP), 0.45pm by using a Trans-Blot Cell (Bio-Rad USA) and was ran at a constant current.of 12 A for 30 minutes. After the transfer, unbound sites on NCP were blocked by incubation with 1 % blocking reagent (Roche Diagnostic) for one hour at room temperature (RT). The NCP was then washed 5 times at 10 min intervals in washing buffer (Tris- HCL buffered saline-tween20, TBST), cut into strips, followed by incubation overnight at 4°C with primary antibody, which comprised serum samples (1 :25 dilution) from the various categories of individuals. All serum samples were absorbed with RF absorbent (Virion-Serion, Germany) according to the manufacturer's instructions. After another washing step, the strips were probed with the appropriate antibody conjugates, namely monoclonal anti-human IgM conjugated to horseradish peroxidase (Zymed, USA) or monoclonal anti-human IgA (Zymed, USA) conjugated to horseradish peroxidase, at a pre-optimised dilution of 1 :4000 and 1 :2000, respectively. After 1 hour incubation, the strips were washed again and subsequently developed by using BM chemiluminescence blotting^' substrate (Roche Diagnosrtic, Germany) and Kodak films (Kodak, USA). The molecular weight of proteins was determined by reference to commercially available standards (Precision Plus Protein standards and unstained marker, BioRad, USA).

' . ^{' ■ ■}:^{' ■■ ■ ' '■■' '■' ■} . ^" . ^■.^{' ■} · Isoelectric focusing

For p/-based protein separation, the 3100 OFFGEL Fractionator and the OFFGEL Kit pH 3-10 (both from Agilent Technologies) with a 24-well setup was used according to the manufacturer's protocol. Fifteen min prior to sample loading, a 24-cm-long IPG gel strip with a linear pH gradient ranging from 3 to 10 was rehydrated in the assembled device with 40 μΙ of rehydration buffer per well. Bromophenol blue was added to rehydration solution at a concentration of 0.002%. Total protein ESA of 500 pg was diluted in focusing buffer to a final volume of 3.6 ml and 150 μΙ sample volume loaded into each well. The sample was focused with a maximum current of 50 μΑ, with a maximum power of 200 mW and voltage ranging from 400 to 4000 V until 64 kVh was reached after approximately 30 hrs. Subsequently, each fraction of 100 ml was collected and electrophoresed on a ^' 12% SDS-PAGE gel and stained with MS compatible silver stain,. Concurrently immunoblotting of these fractions was . performed to confirm the location of the antigenic band with respect to the SDS-PAGE gel. In-Gel digestion and sample clean up

The 10, 12, 20 and 30 kDa protein bands were manually excised from the above silver-stained gel. In-gel digestion of the excised protein bands was performed. First, the gel was destained by adding 100 mM of sodium thiosulphate and 30 mM of potassium ferricyanide in a ratio of 1 :1 for 20 minutes. Then the supernatant was removed and replaced by 100 μΙ of 200 mM ammonium bicarbonate for 20 minutes at room temperature. Destained protein bands were then incubated for 15 minutes in 50 μΙ of acetonitrile and the supernatant was removed, followed by rehydration in 25 μΙ of 25 mM ammonium bicarbonate for 10 minutes. Again, the supernatant was removed and air dried for 1 hour. It was enzymatically digested by re-suspending in 10 ng/μΙ trypsin on ice for 5 minute; the gel pieces were then immersed in 25 μΙ of 25 mM ammonium bicarbonate and incubated overnight at 37°C. Next, acetonitrile was added and the mixture was left for 20 minutes at room temperature. The supernatant which contained the digested protein samples was transferred to fresh Eppendorf tubes and cleaned up by ZipTipcis pipette tips (Milipore).

Mass spectrometry and database analysis

Mass spectrometry analysis of the selected samples were performed using a MALDI-TOF/TOF mass spectrometer. MASCOT search engine (version 2.1 ; Matrix Science) was used to search all of the tandem mass spectra. Analyses were conducted using datasets from ToxoDB, the genome database for the genus Toxoplasma (http://toxodb.Org/common/downloads/release-5.2/Tqondii/).

Results

In both IgA and IgM blots, there were four antigenic bands which reacted well with sera from Group I, namely 10 kDa (Figs. 1 and 2), 12, 20 and 30 kDa (Figs. 2, 3 and 4). Table 1 shows the sensitivities and specificities of each band towards the different categories of sera samples. When IgM and IgA blots were combined, 10 and 12 kDa bands gave sensitivities of 80% and 96% respectively among serum samples from probable active infection (Group I); specificities of 90% and 93 % ^' respectively when tested with sera from chronic infection . (Group II); and .specificities of 97% and 90% respectively when tested with sera from healthy- normal individuals (Group III) and individuals having other infections (Group IV) (Table 1 ). Among Group I serum samples, 20 and 30 kDa bands were seen in IgM immunoblots at a rate of 54/ 75 (72 %) and 43/75 (57 %); and in IgA immunoblots the two bands were detected at a rate of 33/75 (44%) and 40/75 (53%) respectively.

Combination of 12, 20 and 30 kDa bands showed 100% sensitivity in IgM blots in the group of sera that had evidence of , circulating antigen (table 2). Immunoblotting performed with the control antigen did not produce any reactive bands. The isoelectric focusing analysis showed that the p/ range of each 10, 12, 20 and 30 kDa bands were as follows : 5.19- 5.45., 7-7.55, 5.45- 5.71 , 6.24-6.50.

From the results of the MALDI-TOF-TOF and database analysis, the four proteins of molecular weights 10, 12, 20 and 30 kDa was identified as follows respectively: (a) ubiquitin / ribosomal protein CEP52 fusion protein or polyubiquitin (putative), (b) thioredoxin (putative), (c) microneme protein 10 or dense granule protein 7, and (d) phosphoglycerate mutase 1 (putative) or phosphoglycerate mutase (putative). Table 3 shows the details of each protein based on the mass-spectrometry analysis.

The sequence listing accompanying this description shows the amino acid sequences of the four proteins and the nucleotide sequence of their DNA coding sequences. In the sequence listing, the nucleotide coding sequence of the 10 kDa protein is denoted as SEQ ID NO. 1 and its amino acid sequence is SEQ ID NO. 2; alternatively, its nucleotide coding sequence may be SEQ ID NO. 9 with an amino acid sequence of SEQ ID NO. 10. The nucleotide coding sequence of the 12 kDa protein is denoted as SEQ ID NO. 3 and its amino acid sequence is SEQ ID NO. 4. The nucleotide coding sequence of the 20 kDa protein is denoted as SEQ ID NO. 5 and its amino acid sequence is SEQ ID NO-. 6; alternatively, its nucleotide coding sequence may be SEQ ID N0..1 , v» ith an amino acid sequence of SEQ ID NO.12. The nucleotide coding sequence of the 30 kDa protein is denoted as SEQ ID NO. 7 and its amino acid sequence is SEQ ID NO. 8; alternatively, its nucleotide coding sequence may be SEQ ID NO. 13, with an amino acid sequence of SEQ ID NO. 14.

Table 1 : Summary of the reactivities of the four antigenic ESA bands when probed with anti-human IgM and IgA antibodies

*Sensitivity Secondary peroxidase conjugated

Antigenic

and antibodies

proteins

**specificity IgM ^■ IgA IgM & IgA

Sensitivity to

24/30, 80% 24/30, 80 % 24/30, 80% lgM+ lgG+/-

Specificity for .

10 kDa 2/30, 93.3% 3/30, 90% 3/30, 90%

IgM— lgG+

Specificity for

1/30, 96.7 % 1/30, 96.7 % 1/30, 96.7 % IgM- IgG-

Sensitivity to

60/75, 80 % 48 /75, 64% 72/75, 96% lgM+ lgG+/-

Specificity for

12 kDa 3/70, 95.7% 1/30, 97% 2/30, 93.3%

IgM- lgG+

Specificity for

2/50, 96% 0/20, 100% 2/20, 90% IgM- lgG-

Sensitivity to

54/75, 72 % 33/75, 44% 59/75, 78.7% lgM+ lgG+/-

Specificity for

20 kDa 19/70, 87% 10/30, 66.7% 10/30, 66.7%

IgM- lgG+

Specificity for

8/50, 84% 6/20, 70% 6/20, 70% IgM— IgG-

Sensitivity to

43/75, 57 % 40/75, 53% 56/75, 74.7% lgM+ lgG+7-

Specificity for

30 kDa 7/70, 90% 7/30, 76.7% 7/30, 76.7%

IgM- lgG+

Specificity for

4/50, 92% 3/20, 85% 3/20, 85% IgM- IgG- - ^{■ ' '} .19 . . ^{' '}

Table 2: Summary of the results of the ESA bands when incubated with serum samples which were positive for circulating antigen, and probed with anti-human IgM and IgA antibodies

MW of Positive

Positive rates Total Positive antigenic rates in IgA

in IgM blots rates

bands blots

7/10 (70%)

10 kDa 7/10 (70%) 7/10 (70%

20/21 (95%)

12 kDa 19/21 (91 %) 14/21 (67%)

18/21 (86%)

20 kDa 18/21 (86 %) 8 /21 (38%)

17/21 (81 %)

30 kDa 14/21 (67 %) 11 /21 (52%)

Combination

of 12, 20 30 21/21 (100%) 16/21 (76.2%) 21/21 (100%)

kDa

Tabie .3: Mass spectrometric results of ESA antigenic proteins analysed using MASCOT search engine and ToxoDB database

Antigenic Protein ID Accession pi Theoretical Search proteins Number range Mass (Da) Score/ coverage

10 kDa 1. T. gondii_ME49 TGME49_ 089750 9.84 14961 107 product: ubiquitin / NCBI: (score) ribosomal protein XM_002368359

CEP52 fusion

protein

(SEQ ID NO. 2)

2. T. gondii_ME49

product:

TGME49_0 9820 104 polyubiquitih,

putative (SEQ ID

, NO. 10)

12 kDa T. gondii_ME49 TGME49_093870 4.78 12036 53 product: NCBI: (coverage) Thioredoxin XM_002370147

putative

(SEQ ID NO. 4)

20 kDa 1. T. gondii_ME49 TGME49_050710 5.74 23213 77 product : NCBI: (score) microneme protein XM_002367312

10 (SEQ ID NO. 6)

2. T. gondii_ME49

product: dense

granule protein 7 70 (SEQ ID NO. 12) TGME49_003310

30 kDa 1. T. gondii JAE49 TGME49_097060 6.46 30047 113 product:

NCBI: (score) phosphoglycerate

mutase 1 , putative XM_002371095

(SEQ ID NO. 8)

2. T, gondii _GT1

product=phosphog

lycerate mutase, 113

. putative TGGT1._097170

(SEQ ID NO. 14)

Claims

I/We claim:

1 . A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced b or present in a human or animal body in response to Toxoplasma sp. infection, comprising the steps of:

firstly, contacting a biomarker with the biological sample for a time and under conditions sufficient to form a complex with biological products in the biological sample; .

then, detecting the presence or absence of said complex in the biological sample; and

then optionally, analysing the result of said detection by comparing - the result with. a comparison sample or a biological reference sample or both;

wherein said biomarker is characterized by, derived from or made using:

an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO, 12, SEQ ID NO. 14, or any combination thereof; or

an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO. 6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 10, antibodies to SEQ ID NO. 12, antibodies to SEQ ID NO. 14, antibodies to SEQ ID NO. 1 , antibodies to SEQ ID NO. 3, antibodies to SEQ ID NO. 5, antibodies to SEQ ID NO. 7 antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO: 1 1 , antibodies to SEQ ID NO. 13, or any combination thereof; or

a nucleotide sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO-, t, SEQ ID NO. 3,^' SECHD NO. 5, SEQ ID NO..7, SEQ ID NO. 9, SEQ ID NO.- 1 1 , SEQ ID NO. 13, or any combination thereof; or

any combination thereof; and

wherein said method may optionally be carried out using a detection kit.

A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection according to claim 1 , wherein the Toxoplasma sp. is Toxoplasma gondii.

A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in. response to Toxoplasma sp. infection according to claim 1 , wherein the biological sample is whole blood or blood serum or cerebrospinal fluid or eye fluid from a human or animal.

A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection according to claim 1 , wherein the substances produced by or present in Toxoplasma sp. are polypeptides.

A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection according to claim 4, wherein the polypeptides comprise an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, or SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof. A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp.. infection according to claim 1 , wherein the substances produced by or present in a human or animal body in response to Toxoplasma sp. infection are antibodies. .

A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection according to claim 6, wherein the antibodies are ani\-Toxoplasma IgM or IgA or IgG or any combinations thereof.

A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection, according to claim 1 , wherein the biomarker is characterized by, derived from or made using an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof and wherein the biological products in the biological sample are antibodies produced in a human or animal body in response to Toxoplasma sp. infection. .

A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection, according to claim 1 , wherein the biomarker is characterized by, derived from or made, using an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO. 6, antibodies-to SEQ ID NO. 8, antibodies to SEQ ID- NO: 10, antibodies to SEQ ID NO. 12, antibodies to SEQ ID NO. 14, antibodies to SEQ ID NO. 1, antibodies to SEQ ID- NO^'. 3, antibodies to SEQ ID NO: 5, antibodies to SEQ ID NO. 7, antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO. 1 1 , antibodies to SEQ ID NO. 13, or any combination thereof and wherein the biological products in the biological sample are Toxoplasma sp. antigens.

10. A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection, according to claim 1 , wherein the biomarker is characterized by, derived from or made using a nucleotide sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 1 , SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 11 , SEQ ID NO. 13, or any combination thereof and wherein the biological products in the biological sample are antibodies produced in a human or animal body in response to Toxoplasma sp. infection.

1 1. A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection, according to claim 1 , wherein the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof, includes native or recombinant forms.

12. A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection, according to claim 1 , wherein the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID

^' .^' NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8,. SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof, includes antigenic · epitopes thereof.

13. A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection, according to claim 1 , wherein the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of the antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ _. ID NO. 6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 10, antibodies to SEQ ID NO. 12, antibodies to SEQ ID NO. 14, antibodies to- SEQ ID NO. 1 , antibodies to SEQ ID NO. 3, antibodies to SEQ ID NO. 5, antibodies to SEQ ID NO. 7, antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO. 1 1 , antibodies to SEQ ID NO.13, or any combination thereof includes any form such as monoclonal, recombinant or polyclonal antibodies.

14. A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection, according to claim 1 , wherein the nucleotide sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 1 , SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID ^' NO. 11 , SEQ ID NO. 13, or any combination thereof, ma be native or recombinant forms.

15. A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in _. a human or animal body in response to Toxoplasma sp. infection,^' according to claim 1 , wherein the detection kit for carrying out said method comprises the biomarker, a detection reagent for detecting said complex in the biological sample, optionally the comparison sample, and optionally the biological reference sample, wherein said biomarker, detection reagent, comparison sample and biological .reference sample are present in amounts sufficient to perform said method.

16: A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection, according to claim 1 , wherein the comparison sample comprises substances able to specifically bind to the biomarker.

17. A method for detecting, in a biological sample, substances produced by or present in Toxoplasma sp., or substances produced by or present in a human or animal body in response to Toxoplasma sp. infection, according to claim 1 , wherein the biological reference sample is a sample that lacks substances able to bind to the biomarker.

18. A medicament for management or prophylaxis of Toxoplasma sp. infection ^' or a disease resulting therefrom in humans or animals, having a medicinal compound characterized by:

an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO, 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof; or

an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO. 6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 10, antibodies to SEQ ID NO. 12, antibodies to SEQ ID NO. 14, antibodies to SEQ ID NO. 1 , antibodies to SEQ ID NO. 3, antibodies to SEQ ID NO. 5, antibodies to SEQ ID NO. 7 antibodies to SEQ ID NO. 9, antibodies _. to SEQ ID NO. 1 1 , antibodies to SEQ ID NO. 13, or any combination thereof; or

a nucleotide sequence, substantially identical, in part or in whole, to a · • sequence selected- from the group consisting of SEQ 1D NO. 1 , SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO..7, SEQ ID NO. 9, SEQ ID NO. 11 , SEQ ID NO. 13, or any combination thereof; or

any combination thereof;

wherein the medicinal compound may be present alone or in combination with other medicinal substances, and wherein said medicament may further include an acceptable carrier, diluent, or excipient.

19. A medicament for management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals according to claim 18,

• wherein said disease is toxoplasmosis or complications thereof.

20. A medicament for management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals according to claim 18, wherein the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof, includes native, or recombinant forms.

21. A medicament for management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals according to claim 18, wherein the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO; 14, or any combination thereof, includes antigenic epitopes thereof.

22. A medicament for management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals according to claim 18, wherein the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of the antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO. 6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 10, antibodies- to SEQ ID NQ. 12, antibodies to SEQ ID NO. 14,-antibodies to SEQ ID NO 1 , antibodies to SEQ ID NO. 3, antibodies to SEQ ID NO. 5 antibodies to SEQ ID NO. 7, antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO. 11 , antibodies to SEQ ID NO. 13, or any combination thereof includes any form such as monoclonal, recombinant or polyclonal antibodies.

23. A product, or a compound for use in the manufacture of a product, for diagnosis, management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals, wherein the compound is derived from, made using or characterized by:

an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof; or

an amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of antibodies to SEQ ID NO.

2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO. 6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 10, antibodies to SEQ ID NO. 12, antibodies to SEQ ID NO. 14, antibodies to SEQ ID NO. 1 , antibodies to SEQ ID NO. 3, antibodies to SEQ ID NO. 5, antibodies to SEQ ID NO. 7 antibodies to SEQ ID NO. 9, antibodies to SEQ ID NO. 11 , antibodies to SEQ ID NO. 13, or any combination thereof; or

a nucleotide sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 1 , SEQ ID NO.

3, SEQ ID NO. 5, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 11 , SEQ ID NO. 13, or any combination thereof; or

any combination thereof;

24. A product, or a compound for use in the manufacture of a product, for diagnosis, management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals according to claim 23, wherein said disease is toxoplasmosis or complications thereof. ^' 25. A product, or a compound for use in the manufacture of a product, for diagnosis, management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals according to claim 23, wherein the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof, includes native or recombinant forms.

26. A product, or a compound for use in the manufacture of a product, for diagnosis, management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals according to claim 23, wherein the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of SEQ ID NO. 2, SEQ ID

NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, or any combination thereof includes antigenic epitopes thereof.

27. A product, or a compound for use in the manufacture of a product, for diagnosis, management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals according to claim 23, wherein the amino acid sequence substantially identical, in part or in whole, to a sequence selected from the group consisting of the antibodies to SEQ ID NO. 2, antibodies to SEQ ID NO. 4, antibodies to SEQ ID NO. 6, antibodies to SEQ ID NO. 8, antibodies to SEQ ID NO. 0, antibodies to SEQ ID NO. 12, antibodies to SEQ ID NO. 14, antibodies to SEQ ID NO. 1 , antibodies to SEQ ID NO. 3, antibodies to SEQ ID NO. 5 antibodies to SEQ ID NO. 7, antibodies to SEQ ID NO. 9, antibodies to SEQ ID. NO: 11 , antibodies to SEQ ID NO.13, or any combination thereof includes any form such as monoclonal, recombinant or polyclonal antibodies.

28. A product, or a compound for use in the manufacture of a product, for diagnosis, management or prophylaxis of Toxoplasma sp. infection or a disease resulting therefrom in humans or animals according to. claim 23, wherein said product includes gel electrophoresis bands, or electrophoresis bands transferred onto a membrane, or any combination thereof.