Serological Study on Cytomegalovirus and Toxoplasma Gondii in Thalassemia Major Patients of Yazd, Iran.
Minoosh MoghimiMasoud DoostiHA Vahedian-ArdakaniAlireza TalebiMahvash AkhavanghalibafAtabak NajafiM M AminorroayaShahrooz YazdaniMohammad ShayestehpourH A BahramiF Khodayari
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Abstract:
Beta-thalassemia patients receive blood products from blood transfusion centers repeatedly. Blood transfusion can transmit Cytomegalovirus (CMV) and Toxoplasma gondii. The aim of this study was serological evaluation of these two infectious agents in thalassemia patients.In a cross-sectional study, the enzyme-linked immunosorbent assay (ELISA) testing was performed to detect IgM and IgG antibodies against CMV and Toxoplasma gondii in 96 thalassemia patients (under 18 years) and 144 healthy people. Data were analyzed by SPSS software and Chi-square test.A significant difference was observed in CMVIgM antibody levels between test groups in women (p<0.05). The prevalence of CMV IgM, CMV IgG, Toxo-IgG, and Toxo IgM antibodies in thalassemia patients were 5.2%, 95.9%, 16%, and 0%, respectively.In all thalassemia patients, Cytomegalovirus IgG is higher than healthy people. In addition, CMV IgM antibodies are higher in female patients. Antibody screening (IgM) on blood products for detecting Cytomegalovirus is necessary, but for Toxoplasma gondii is not necessary in the Yazd transfusion center.Keywords:
Cytomegalovirus
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Beta-thalassemia patients receive blood products from blood transfusion centers repeatedly. Blood transfusion can transmit Cytomegalovirus (CMV) and Toxoplasma gondii. The aim of this study was serological evaluation of these two infectious agents in thalassemia patients.In a cross-sectional study, the enzyme-linked immunosorbent assay (ELISA) testing was performed to detect IgM and IgG antibodies against CMV and Toxoplasma gondii in 96 thalassemia patients (under 18 years) and 144 healthy people. Data were analyzed by SPSS software and Chi-square test.A significant difference was observed in CMVIgM antibody levels between test groups in women (p<0.05). The prevalence of CMV IgM, CMV IgG, Toxo-IgG, and Toxo IgM antibodies in thalassemia patients were 5.2%, 95.9%, 16%, and 0%, respectively.In all thalassemia patients, Cytomegalovirus IgG is higher than healthy people. In addition, CMV IgM antibodies are higher in female patients. Antibody screening (IgM) on blood products for detecting Cytomegalovirus is necessary, but for Toxoplasma gondii is not necessary in the Yazd transfusion center.
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An assay measuring the avidity of T. gondii-specific IgG is a useful serological indicator of toxoplasmosis which in many cases allows on the basis of single serum sample testing to confirm or exclude acute infection. IgG antibodies against T. gondii produced in the recent primary infection are of low-avidity while IgG antibodies in the chronic toxoplasmosis are of high-avidity. In the present study 80 sera: 47 sera of patients with suspected acute toxoplasmosis, 23 sera of pregnant women and 10 sera of healthy blood donors, were evaluated for the presence of IgM, IgA and IgG antibodies and for the avidity of IgG. Among the 80 tested sera IgM antibodies were present in 34 (42.5%) cases of which 22 (64.7%) showed low-avidity of IgG and the presence of IgA antibodies confirming acute toxoplasmosis. In the rest of these 34 sera three of them showed low avidity index and nine other high avidity in spite of the presence of IgM antibodies. In these 12 sera IgA antibodies were not present. In 46 (57.5%) examined sera IgM or IgA antibodies were not detectable and the present the IgG antibodies showed high-avidity what is characteristic of chronic infection. An assay for evaluation of avidity of IgG antibodies specific for T. gondii is valuable in complementing existing tests and in many cases it has decisive value for interpretation of results.
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The persistence, in some subjects, of specific IgM antibodies to Toxoplasma gondii for several months after the acute phase of infection has complicated the interpretation of serological test results for toxoplasmosis. Several reports have emphasized the value of the detection of Toxoplasma-specific IgA antibodies for the diagnosis of acute toxoplasmosis. In this article, we report the follow-up profiles of Toxoplasma-specific IgM and IgA antibodies in serum samples obtained from 12 patients at various intervals after the onset of the clinical manifestations of infection. IgM antibodies were detected by the indirect immunofluorescence (IIF) test, antibody capture enzyme-linked immunosorbent assay (cELISA) and enzyme-mediated chemilluminescent technique (CmL). IgA antibodies were quantified by the direct ELISA (dELISA) and cELISA procedures. As defined by the manufacturer of the cELISA test for IgA used, most patients with acute toxoplasmosis have antibody levels > 40 arbritary units per ml (AU/ml). At values > 40 AU/ml, the cELISA for IgA detected significant antibody levels for a shorter time than the other techniques used for IgM and IgA detection. However, IgA levels < or = 40 AU/ml do not exclude the possibility of acute toxoplasmosis since such levels can be reached very soon after infection with T. gondii. The results obtained in the present study show that the serological diagnosis of acute toxoplasmosis may not be such an easy task. Our data suggest that use of the IgA-cELISA concomitantly with IgM antibody screening could permit, in some circumstances, a more efficient diagnosis of acute acquired toxoplasmosis.
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Toxoplasma gondii causes congenital toxoplasmosis in newborns resulting with fetal anomalies. Determining the initiation time of infection is very important for pregnant women and current serological assays have drawbacks in distinguishing the recently acute toxoplasmosis. Diagnosis of recently acute infection may be improved by using stage specific antigens in serological assays. In the present study, the diagnostic value of sporozoite specific SporoSAG, bradyzoite specific BAG1 proteins and GRA1 protein expressed by all forms of the parasite have been evaluated ELISA using sera systematically collected from mice administered orally with tissue cyst and oocysts. The anti-SporoSAG IgM antibodies in sera obtained from mice infected with oocysts peaked significantly at days 1, 10, and 15 (P<0.01). The anti-BAG1 IgM antibodies in sera obtained from mice infected with tissue cysts peaked significantly at days 15, 40, and 120 (P<0.05). The anti-GRA1 IgM antibodies in sera obtained from mice infected with oocysts peaked significantly at days 2, 10, and 40 (P<0.01). The anti-GRA1 IgM antibodies in sera obtained from mice infected with tissue cysts peaked significantly only at day 40 (P<0.05). The anti-SporoSAG, anti-BAG1, and anti-GRA1 IgG titers of mice showed significant increases at day 40 (P<0.05) and decrement started for only anti-GRA1 IgG at day 120. The presence of anti-SporoSAG IgM and IgG antibodies can be interpreted as recently acute infection between days 10-40 because IgM decreases at day 40. Similarly, presence of anti-BAG1 IgM and absence of IgG can be evaluated as a recently acute infection that occurred 40 days before because IgG peaks at day 40. A peak in anti-GRA1 antibody level at first testing and reduction in consecutive sample can be considered as an infection approximately around day 40 or prior. Overall, recombinant SporoSAG, BAG1 and GRA1 proteins can be accepted as valuable diagnostic markers of recently acute toxoplasmosis.
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A 4-layer modification of ELISA for the determination of toxoplasma antibodies is described. In 103 Finnish blood donors 27 had antibodies against Toxoplasma gondii. One donor had IgM antibodies and IgA antibodies were found in 9. In patients with acute toxoplasmosis a vigorous IgG antibody response resulted in high antibody levels soon after infection, declining gradually to mean adult levels in approximately 2 yr. IgM antibodies appeared during the earliest phases of infection and disappeared as early as in 1 or 2 months in some cases and in most cases by the 6th month after infection. An IgA antibody response was also always seen. It was slower than the IgM response and could therefore be used to improve the laboratory diagnosis especially in cases where IgM antibodies had already disappeared and no further increase in IgG antibodies could be detected.
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Immunoglobulin M (IgM) and IgA antibodies against the major surface protein of Toxoplasma gondii were determined in a total of 195 human sera and five human cerebrospinal fluids by using a P30 membrane extract and the immunoblot technique. By using two different T. gondii strains (RH and BK) simultaneously as antigens, we were able to demonstrate diagnostically important strain-specific human antibody responses in 4.5% of the samples tested. A comparison of the immunoblot technique with an IgM immunocapture enzyme-linked immunosorbent assay demonstrated that the IgM and IgA immunoblot seems to be of advantage in the diagnosis of acute toxoplasmosis in certain groups of patients, especially in the diagnosis of cerebral toxoplasmosis in patients with AIDS. The immunoblot technique described is easy to perform and might be useful as an additional serological assay for routine diagnosis of T. gondii infections.
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The diagnosis of Toxoplasma gondii infection is currently based on immunological tests, but tests for IgM and IgG antibodies alone are often insufficient to estimate the risk of active disease, especially during pregnancy and in immunodeficient patients. Classically the study of anti-toxoplasma immunity involves titration of IgG antibodies, which reflect immunity to the parasite, and IgM antibodies which of present, reveal acute infection. However, technical advances have shown the limitations of these tests as tests for IgM can be positive because of residual specific IgM or even in subjects free of acute infection due to the existence of natural interfering IgM. In addition, IgM can be absent in children with congenital toxoplasmosis or subjects with secondary reactivation. The purpose of our study was to evaluated of IgA antibodies to T. gondii in serum samples which were positive in screening test. Our results confirm the diagnostic value of testing for anti-toxoplasma IgA antibodies. These antibodies are absent in uninfected subjects and are detected rapidly after primary infection. The determination of IgA complements IgM determination for the diagnosis of toxoplasmosis.
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