New recombinant chimeric antigens, P35-MAG1, MIC1-ROP1, and MAG1-ROP1, for the serodiagnosis of human toxoplasmosis.

The aim of the study was to evaluate the usefulness of 3 chimeric Toxoplasma gondii antigens, P35-MAG1, MIC1-ROP1 and MAG1-ROP1, in the serodiagnosis of an acute toxoplasmosis in humans. Proteins were produced as fusion proteins containing His tags ends and then further purified by metal affinity chromatography. Their application for the diagnosis of recently acquired T. gondii infection was tested in IgG and IgM enzyme-linked immunosorbent assays (ELISAs). At 100%, 77.3%, and 86.4%, respectively, the reactivity of the IgG ELISA using P35-MAG1, MIC1-ROP1, and MAG1-ROP1 for sera from patients where acute toxoplasmosis was suspected was significantly higher than for the samples from people with a chronic infection, at 26.2%, 36.1%, and 32.8%, respectively. Moreover, P35-MAG1, MIC1-ROP1, and MAG1-ROP1 detected IgM antibodies with a reactivity at 81.8%, 72.7%, and 59.1%, respectively. The results presented in the article show that, particularly, P35-MAG1 may be useful in the preliminary detection of recent T. gondii infection.

A new human IgG avidity test, using mixtures of recombinant antigens (rROP1, rSAG2, rGRA6), for the diagnosis of difficult-to-identify phases of toxoplasmosis.

The preliminary diagnostic utility of two mixtures of Toxoplasma gondii recombinant antigens (rROP1+rSAG2 and rROP1+rGRA6) in IgG ELISA and IgG avidity test has been evaluated. A total of 173 serum samples from patients with toxoplasmosis and seronegative people were examined. The sensitivity of IgG ELISA for rROP1+rSAG2 and rROP1+rGRA6 was 91.1% and 76.7%, respectively, while the reactivity for sera from patients where acute toxoplasmosis was suspected was higher, at 100% and 95.4%, respectively, than for people with chronic infection, at 88.2% and 70.6%. In this study a different trend in avidity maturation of IgG antibodies for two mixtures of proteins in comparison with native antigen was observed. The results suggest that a new IgG avidity test using the mixtures of recombinant antigens may be useful for the diagnosis of difficult-to-identify phases of toxoplasmosis. For this reason, selected mixtures after the additional tests on groups of sera with well-defined dates of infection could be used as a better alternative to the native antigens of the parasite in the serodiagnosis of human T. gondii infection.

Epidemiological study of Toxoplasma gondii infection among cattle in Northern Poland.

Toxoplasmosis, caused by Toxoplasma gondii, is a significant disease in livestock and humans. Because of medical and veterinary importance it is essential to study the prevalence of T. gondii infection among human and animals in various parts of the word. In this study, 4033 cattle from eight provinces of Northern Poland (belonging to 190 herds) were tested for IgG antibodies against T. gondii by an in-house ELISA technique based on native Toxoplasma lysate antigen. The diagnostic sensitivity of test used in this study was 96.3%, and specificity was 98% for the group of 77 cattle sera (27 seropositive and 50 seronegative) previously characterized with the use of agglutination and immunofluorescence methods. A 127 (3.15%) out of all tested animals belonging to 72 (37.9%) out of 190 herds were founded as positive. Furthermore, our results showed that the way of feeding and farming, the size of the herd and the age of animals have the influence on the prevalence of toxoplasmosis among cattle. The percentage of infected cattle was the highest for old animals which belongs to the small herds with the traditional way of farming. These results indicate that T. gondii infection in cattle from Northern Poland is relatively low and consumption of beef and milk can be regarded as a poor source of infection for humans.

MIC1-MAG1-SAG1 chimeric protein, a most effective antigen for detection of human toxoplasmosis.

This study describes a Toxoplasma gondii IgG enzyme-linked immunosorbent assay based on a new chimeric antigen containing three immunodominant regions from the MIC1, MAG1, and SAG1 proteins of the parasite and shows that this test is useful for diagnostic purposes and may replace the lysed and whole-cell antigens.

A new MIC1-MAG1 recombinant chimeric antigen can be used instead of the Toxoplasma gondii lysate antigen in serodiagnosis of human toxoplasmosis.

This study presents an evaluation of the MIC1 (microneme protein 1)-MAG1 (matrix antigen 1) Toxoplasma gondii recombinant chimeric antigen for the serodiagnosis of human toxoplasmosis for the first time. The recombinant MIC1-MAG1 antigen was obtained as a fusion protein containing His tags at the N- and C-terminal ends using an Escherichia coli expression system. After purification by metal affinity chromatography, the chimeric protein was tested for usefulness in an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-T. gondii immunoglobulin G (IgG). One hundred ten sera from patients at different stages of infection and 40 sera from seronegative patients were examined. The results obtained for the MIC1-MAG1 chimeric antigen were compared with those of IgG ELISAs using a Toxoplasma lysate antigen (TLA), a combination of recombinant antigens (rMIC1ex2-rMAG1) and single recombinant proteins (rMIC1ex2 and rMAG1). The sensitivity of the IgG ELISA calculated from all of the positive serum samples was similar for the MIC1-MAG1 chimeric antigen (90.8%) and the TLA (91.8%), whereas the sensitivities of the other antigenic samples used were definitely lower, at 69.1% for the mixture of antigens, 75.5% for the rMIC1ex2, and 60% for rMAG1. This study demonstrates that the MIC1-MAG1 recombinant chimeric antigen can be used instead of the TLA in the serodiagnosis of human toxoplasmosis.

Toxoplasma gondii: usefulness of ROP1 recombinant antigen in an immunoglobulin G avidity assay for diagnosis of acute toxoplasmosis in humans.

The results present in this study suggest that the Toxoplasma gondii recombinant ROP1 antigen in an IgG avidity test can be useful for detection of acute stage of infection. Specific antibodies of low avidity were detected in most of the sera from individuals with acute toxoplasmosis, while the absence or specific antibodies of high avidity were detected in sera from patients with chronic infection.