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      Detection of Toxoplasma gondii bradyzoite genes in the peripheral blood mononuclear cells among patients with toxoplasmic chorioretinitis

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          Abstract

          Background

          Toxoplasmic chorioretinitis may occur as a result of acquired toxoplasmosis or reactivated congenital toxoplasmosis. In this study, Toxoplasma gondii bradyzoite genes along with the B1 gene were evaluated to detect T. gondii DNA in serum and peripheral blood mononuclear cells (PBMCs) of patients with toxoplasmic chorioretinitis.

          Methods

          Blood samples were collected from 10 patients (7 cases of active chorioretinal lesions and 3 cases of old chorioretinal scars). The genomic DNA was extracted from the patients’ serum and PBMCs and a polymerase chain reaction (PCR) assay was performed using bradyzoite genes along with B1. The subjects were also evaluated in terms of the T. gondii antibodies.

          Results

          The PCR results were positive in four of seven patients (57.1%) with active ocular toxoplasmosis lesions. In three patients (42.8%), the PCR results were positive for MAG-1 and SAG-4 and in one patient (14.3%) the PCR results were only positive for the B1 gene. The PCR results were positive only in the PBMCs, whereas they were negative in the serum samples. Two patients with positive PCR results showed high Toxoplasma immunoglobulin G (IgG) antibody titres. However, none of the patients showed positive Toxoplasma IgM antibodies.

          Conclusions

          The PBMCs are suitable for evaluating toxoplasmic chorioretinitis. The present results showed that PCR with bradyzoite genes is useful in the diagnosis of toxoplasmic chorioretinitis in PBMCs.

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          Most cited references25

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          A follow-up study of Toxoplasma gondii infection in southern Brazil.

          To understand better the natural history of ocular toxoplasmosis by reexamining a well-characterized population in Southern Brazil. Ophthalmological examination and serologic tests for Toxoplasma gondii infection were performed in 1997 on 383 individuals who had undergone the same evaluation in 1990. Of 109 seronegative subjects in 1990, 21 (19.3%) became seropositive by 1997, and 2 (1.5% of previously seronegative patients; 9.5% of those known to have seroconverted) developed ocular toxoplasmosis. Seroconversion occurred more frequently in individuals under 17 years of age (16 of 46 patients, 34.8%) than in those greater than 17 years of age (5 of 63 patients, 7.9%; p = 0.002). Of 131 seropositive individuals who did not have ocular lesions in 1990, 11 (8.3%) had typical toxoplasmic lesions in 1997. Of the 13 individuals with non-specific hyperpigmented small retinal lesions in 1990, 3 (23%) presented with typical lesions in 1997. Acquired T. gondii infection can result in late development of ocular lesions. Small, non-specific hyperpigmented retinal lesions may represent sites of T. gondii infection in seropositive individuals.
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            Specific, sensitive, and rapid diagnosis of active toxoplasmosis by a loop-mediated isothermal amplification method using blood samples from patients.

            Loop-mediated isothermal amplification (LAMP), a rapid nucleic acid amplification method, was developed for the clinical diagnosis of toxoplasmosis. Three LAMP assays based on the SAG1, SAG2, and B1 genes of Toxoplasma gondii were developed. The sensitivities and specificities of the LAMP assays were evaluated by comparison with the results of conventional nested PCR. The LAMP assays were highly sensitive and had a detection limit of 0.1 tachyzoite, and no cross-reactivity with the DNA of other parasites was observed. Blood was collected from 105 individuals to test the LAMP assays: 40 patients with active toxoplasmosis, 40 negative controls, and 25 patients with other parasitic infections. The SAG2-based LAMP (SAG2-LAMP) had a greater sensitivity (87.5%) than the SAG1-LAMP (80%), B1-LAMP (80%), and nested PCR (62.5%). All the LAMP assays and nested PCR were 100% specific. This is the first report of a study which applied the LAMP method to diagnose toxoplasmosis from human blood samples. Due to its simplicity, sensitivity, and specificity, LAMP is suggested as an appropriate method for routine diagnosis of active toxoplasmosis in humans.
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              Toxoplasmic chorioretinitis in the setting of acute acquired toxoplasmosis.

              Ocular toxoplasmosis is considered to be the most commonly recognized cause of chorioretinitis in the United States. It is commonly believed that the majority of cases of acute toxoplasmic chorioretinitis involving adults in the United States are late sequelae of congenital infection and that the condition is rarely associated with acute postnatally acquired infection. We report here the clinical and serological test findings for 22 adults with acute toxoplasmic chorioretinitis that occurred in the setting of acute postnatally acquired toxoplasmosis. The initial serum specimen from each adult yielded an acute toxoplasmic serological profile, on the basis of the following positive results: 95.5%, Sabin-Feldman dye test [titer of > or = 1:1,024]; 95.5%, IgM ELISA; 90.9%, IgA ELISA; 77.3%, IgE ELISA; 95.5%, IgE immunosorbent agglutination assay; and 86.4%, differential agglutination (AC/HS) test (acute pattern). Detection of IgA or IgE antibodies or an acute pattern in the AC/HS test was particularly helpful in diagnosis for those patients whose ELISA IgM titers at presentation were negative or lowly positive. Thus, acute toxoplasmic chorioretinitis occurring with a recently acquired Toxoplasma gondii infection would appear to be more common in the United States than previously recognized, and a toxoplasmic serological profile is useful in diagnosing this entity.
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                Author and article information

                Contributors
                (View ORCID Profile)
                Journal
                Transactions of The Royal Society of Tropical Medicine and Hygiene
                Oxford University Press (OUP)
                0035-9203
                1878-3503
                April 13 2021
                April 13 2021
                Affiliations
                [1 ]Research Center of Pediatric Infectious Diseases, Institute of Immunology and Infectious Diseases, Iran University of Medical Sciences, Tehran, Iran
                [2 ]Eye Research Center, the five Senses Institute, Iran University of Medical Sciences, Tehran, Iran
                [3 ]Noor Ophthalmology Research Center, Noor Eye Hospital, Tehran, Iran
                [4 ]Antimicrobial Resistance Research Center, Institute of Immunology and Infectious Diseases, Iran University of Medical Sciences, Tehran, Iran
                [5 ]Department of Virology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
                [6 ]Vice Chancellor for Healthcare, Iran University of Medical Sciences, Tehran, Iran
                [7 ]Student Research Committee, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
                Article
                10.1093/trstmh/trab062
                b2b6067f-71e7-4cf1-92b4-650f5cf028f3
                © 2021

                https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model

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