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      Antiparasitic treatment suppresses production and avidity of Toxoplasma gondii-specific antibodies in a murine model of acute infection

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          Infection with Toxoplasma gondii during pregnancy may result in congenital transmission of the parasite. Infection is commonly diagnosed using serological tests for IgG, IgM and IgA antibodies. Avidity of IgG antibodies is used to exclude acute infection. Few studies have investigated the impact of antiparasitic treatment on the production of anti- T. gondii antibody and the avidity of IgG antibodies. We therefore investigated the production of IgG, IgM, and IgA antibodies and IgG avidity in a murine model of acute infection with 10 cysts of T. gondii. All antibody classes increased following infection. Treatment of mice with py rime thamine/ sulfadiazine but not with spiramycin or azithromycin at dosages equivalent to those used in patients resulted in a significant decrease in the concentration of T. gondii-specific IgG and IgM antibodies postinfection. IgG and IgM antibody decreases were paralleled by a significant reduction in cyst numbers in brains of mice treated with pyrimethamine/sulfadiazine but not with other drugs. In contrast, treatment with atovaquone did significantly reduce the concentrations of IgM antibodies and resulted in reduced IgG avidity indices. T. gondii-specific DNA was not detected in blood between days 1 and 3. In conclusion, antiparasitic treatment with pyrimethamine/sulfadiazine and atovaquone appears to impact the generation of antibody responses against T. gondii. Future studies will have to determine the specific impact of antiparasitic treatment on antibody responses and the consequences for the management of patients infected with T. gondii.

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          Management of Toxoplasma gondii infection during pregnancy.

          Acute infection with Toxoplasma gondii during pregnancy and its potentially tragic outcome for the fetus and newborn continue to occur in the United States, as well as worldwide, despite the fact that it can be prevented. The infection can be acquired through ingestion of infected, undercooked meat or contaminated food or water. Transmission to the fetus occurs almost solely in women who acquire their primary infection during gestation and can result in visual and hearing loss, mental and psychomotor retardation, seizures, hematological abnormalities, hepatosplenomegaly, or death. Systematic education and serological screening of pregnant women are the most reliable and currently available strategies for the prevention, diagnosis, and early treatment of the infection in the offspring; this is largely because toxoplasmosis in pregnant women most often goes unrecognized. Treatment of the infection in the fetus and infant during the first year of life has been demonstrated to significantly improve the clinical outcome.
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            Laboratory diagnosis of Toxoplasma gondii infection and toxoplasmosis.

            J Montoya (2002)
            For the past 40 years, the Toxoplasma Serology Laboratory at the Palo Alto Medical Foundation Research Institute (TSL-PAMFRI) has been dedicated to the laboratory diagnosis of Toxoplasma gondii infection and toxoplasmosis. TSL-PAMFRI is the "brain child" of Jack S. Remington. Jack's ceaseless devotion to objectivity and uncompromising excellence has made TSL-PAMFRI the Toxoplasma reference laboratory for the Centers for Disease Control and Prevention, the US Food and Drug Administration, and health care providers and clinical laboratories in the United States and other countries. Jack's leadership and vision created, defined, and significantly contributed to the development of laboratory methods for the diagnosis of the infection and diseases caused by T. gondii. A summary of the laboratory tests currently available at TSL-PAMFRI for the diagnosis of infection and disease caused by the parasite is presented here.
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              Comparison of two DNA targets for the diagnosis of Toxoplasmosis by real-time PCR using fluorescence resonance energy transfer hybridization probes

              Background Toxoplasmosis is an infectious disease caused by the parasitic protozoan Toxoplasma gondii. It is endemic worldwide and, depending on the geographic location, 15 to 85% of the human population are asymptomatically infected. Routine diagnosis is based on serology. The parasite has emerged as a major opportunistic pathogen for immunocompromised patients, in whom it can cause life-threatening disease. Moreover, when a pregnant woman develops a primary Toxoplasma gondii infection, the parasite may be transmitted to the fetus and cause serious damnage. For these two subpopulations, a rapid and accurate diagnosis is required to initiate treatment. Serological diagnosis of active infection is unreliable because reactivation is not always accompanied by changes in antibody levels, and the presence of IgM does not necessarily indicate recent infection. Application of quantitative PCR has evolved as a sensitive, specific, and rapid method for the detection of Toxoplasma gondii DNA in amniotic fluid, blood, tissue samples, and cerebrospinal fluid. Methods Two separate, real-time fluorescence PCR assays were designed and evaluated with clinical samples. The first, targeting the 35-fold repeated B1 gene, and a second, targeting a newly described multicopy genomic fragment of Toxoplasma gondii. Amplicons of different intragenic copies were analyzed for sequence heterogeneity. Results Comparative LightCycler experiments were conducted with a dilution series of Toxoplasma gondii genomic DNA, 5 reference strains, and 51 Toxoplasma gondii-positive amniotic fluid samples revealing a 10 to 100-fold higher sensitivity for the PCR assay targeting the newly described 529-bp repeat element of Toxoplasma gondii. Conclusion We have developed a quantitative LightCycler PCR protocol which offer rapid cycling with real-time, sequence-specific detection of amplicons. Results of quantitative PCR demonstrate that the 529-bp repeat element is repeated more than 300-fold in the genome of Toxoplasma gondii. Since individual intragenic copies of the target are conserved on sequence level, the high copy number leads to an ultimate level of analytical sensitivity in routine practice. This newly described 529-bp repeat element should be preferred to less repeated or more divergent target sequences in order to improve the sensitivity of PCR tests for the diagnosis of toxoplasmosis.

                Author and article information

                European Journal of Microbiology and Immunology
                Akadémiai Kiadó, co-published with Springer Science+Business Media B.V., Formerly Kluwer Academic Publishers B.V.
                1 September 2011
                : 1
                : 3
                : 249-255
                [ 1 ] Departamento de Infectologia, Facultad de Medicina y Nutrición, Universidad Juárez del Estado de Durango. Durango, Dgo. México, Avenida Universidad y Fanny Anitua, 34000, Durango, Dgo. Mexico
                [ 2 ] Robert-Koch-Institut, Nordufer 20, D-13353, Berlin, Germany
                [ 3 ] Medical and Scientific Affairs, Roche Molecular Diagnostics, 4300 Hacienda Dr., Pleasanton, CA, 94588, USA
                [ 4 ] Department of Microbiology and Hygiene, Charité Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany
                [ 5 ] Department of Microbiology and Hygiene, Charité Campus Benjamin Franklin, Hindenburgdamm 27, D-12203, Berlin, Germany
                Author notes

                Parts of this work were presented at ECCMID 2008 (abstract P1875).

                [* ] +493084453628, +493084453830, oliver.liesenfeld@ 123456charite.de
                Original Articles

                Medicine,Immunology,Health & Social care,Microbiology & Virology,Infectious disease & Microbiology
                IgG avidity,mice,treatment,antibodies, Toxoplasma gondii infection


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