Diagnosis Of Human Toxoplasmosis Research Articles

Carbon nanospheres dual spectral-overlapped fluorescence quenching lateral flow immunoassay for rapid diagnosis of toxoplasmosis in humans

Toxoplasmosis is a common zoonotic disease caused by a protozoan parasite Toxoplasma gondii (Tox), approximately infecting one-third of human populations worldwide. This study developed the carbon nanospheres (CNPs) based dual spectral-overlapped fluorescence quenching lateral flow immunoassay (CNPs-FQLFIA) for detection of Tox antibodies (ToxAbs). The CNPs have been effectively coupled with Tox antigen (ToxAg), which can completely overlap the excitation and emission spectra of europium nanospheres (EuNPs) and CdSe/ZnS quantum dots (QDs) in testing strips of CNPs-QDs-FQLFIA or CNPs-EuNPs-FQLFIA. The sensitivity of CNPs-EuNPs-FQLFIA or CNPs-QDs-FQLFIA was 4 or 8 IU/mL under natural light readout, or both 4 IU/mL ToxAbs under ultraviolet (UV) light readout by the naked eyes, respectively. The limit of detection (LOD) of two types of CNPs-FQLFIA was both 1 IU/mL ToxAbs under UV light by a dry fluorescence analyzer, but no cross-reaction was found with other antibodies. The intra-assay coefficient variation (CV) of both CNPs-EuNPs-FQLFIA and CNPs-QDs-FQLFIA was less than 8%, while the inter-assay CV was less than 14%, respectively. The correlation coefficient (R2) of CNPs-EuNPs-FQLFIA or CNPs-QDs-FQLFIA to measure the different concentrations of ToxAbs spiked serum samples was 0.99712 and 0.99896, respectively. The CNPs-FQLFIA presented a characteristics of 94.3% sensitivity, 100% specificity and 98% accuracy for detection of ToxAbs in clinical serum samples. In conclusion, CNPs-FQLFIA with EuNPs or QDs fluorescence reporter was an easy, rapid, sensitive, precise and quantitative assay for detecting Tox antibodies in human blood samples.

Assessment of the Performance of Combining Rapid Immunochromatographic and Latex Agglutination Tests in the Diagnosis of Human Toxoplasmosis in Cameroon

Assessment of the Performance of Combining Rapid Immunochromatographic and Latex Agglutination Tests in the Diagnosis of Human Toxoplasmosis in Cameroon

Serological detection of T. gondii infection in humans using an immunochromatographic assay based on dense granule protein 7.

Toxoplasma gondii causes toxoplasmosis, one of the world's most common parasitic diseases. It secretes large amounts of dense granule antigens (TgGRAs), which are crucial to the parasite's survivability. TgGRA7 is found abundantly on the surface of host cells, within the parasitophorous vacuole lumen and membrane, and the host cell cytosol. It stimulates a strong antibody response during acute and chronic infections. While it has been well utilized as an antigen for enzyme-linked immunosorbent assay (ELISA), only one report has documented its efficacy as an antigen for an immunochromatographic test (ICT) in pigs. To date, there is no study yet documenting its use for ICT in human toxoplasmosis. Here, we validated the efficacy of the TgGRA7-ICT we developed by testing 88 human sera. Results were compared with those obtained by ELISA based on TgGRA7, a commercial ELISA, and latex agglutination test (LAT). With high sensitivity, specificity, and kappa values, our TgGRA7-ICT results revealed very good agreement with standard test results. We also found a strong correlation between the relative ICT band intensity and absorbance values in the ELISA. Altogether, our data suggest that the current ICT with TgGRA7 is a reliable test for the diagnosis of human toxoplasmosis, which produced results similar to conventional serological methods. Thus, this can be used as a screening tool for routine testing of toxoplasmosis and a good option for point of care application. The present study also documents the first utilization of TgGRA7 as an antigen for ICT for the serodiagnosis of human toxoplasmosis.

Development of an indirect immunofluorescence technique for the diagnosis of toxoplasmosis in bottlenose dolphins

The diagnosis of toxoplasmosis is often complicated by the lack of specific clinical symptoms or postmortem features, in humans and other animals. The only diagnostic test described so far for the serological diagnosis of Toxoplasma gondii in marine mammals is the modified agglutination test (Dubey et al., Am J Vet Res 48(8):1239-1243, 1987). The development of more sensible and specific immunological techniques requires specific antibodies, which are currently unavailable in the scientific market. Indirect immunofluorescence (IIF) is one of the most widely used methods for the diagnosis of toxoplasmosis in humans (Auer et al., Parasitol Res 12:965-970, 2000). In order to develop and apply this technique to the bottlenose dolphin (Tursiops truncatus), immunoglobulins were firstly purified using ion-exchange chromatography. The purified immunoglobulins were then injected in New Zealand rabbits in order to obtain polyclonal antibodies. These antisera were validated by the IIF technique, using as controls serum samples of dolphins infected by Toxoplasma. The results were visualized using antirabbit IgG labeled with fluorescein. This newly developed and specific serological assay was then tested with the dolphin collection of Loro Parque, Tenerife, Spain (group I), and L'Oceanogràfic of Valencia, Spain (group II). The obtained results in this study showed that none of the dolphins from group 1 were infected by T. gondii and two animals were positive in group 2. Furthermore, we conclude that this study has produced antibodies with high specificity against dolphin immunoglobulins and an IIF method which may be used as immunological diagnostic tools, especially for the serological diagnosis of toxoplasmosis.

Toxoplasmosis: The value of molecular methods in diagnosis compared to conventional methods

Zineb Tlamcani, Zohra Lemkhenete, Badre Eddine Lmimouni Toxoplasmosis is a parasitic infection due to Toxoplasma gondii an obligate intracellular protozoan parasite. It is conssiderate one of the most common parasite worldwide. The contamination of the parasite is generally occurred via conssumption of infected food or water or, undercooked contaminated meat. Toxoplasma gondii infection may lead to serisous illness when the organism is contracted while pregnancy or when it is reactivated in immune-suppressed persons. Diagnosis of toxoplasmosis in humans is elaborated using various techniques such as detection of anti-Toxoplasma antibodies, mouse inoculation, histological revelation of tachyzoites in tissue sections or smears of body fluid, but the detection of Toxoplasma gondii DNA by molecular methods has revolutionized prenatal diagnosis of congenital toxosplasmosis and in immunocompromised patients. In this paper we will discuss the parasite and different methods of diagnosis including the usefulness of molecular methods.

Unspecific reactivity of IgM directed against the low-molecular-weight antigen of Toxoplasma gondii.

During routine serological survey, eight patients (5 pregnant women, 3 grafted patients) were positive for Toxoplasma gondii-specific IgM by enzyme-linked immunoassay but negative by a simultaneously performed immunosorbent agglutination assay. No clinical or biological symptoms of toxoplasmosis were observed later, despite the absence of treatment. Only one IgM-reactive band, which corresponded to the low-molecular-weight antigen of Toxoplasma gondii, was observed by Western blotting of these patients' sera. Dot blotting of lipid extracts of Toxoplasma gondii demonstrated that this reactivity was directed against sphingolipids or ceramides. This IgM positivity, which is unrelated to acute toxoplasmosis, raises strong concerns about the possibility of misleading results of this test in the diagnosis of toxoplasmosis in humans.

Polymerase chain reaction approaches for the detection of Neospora caninum and Toxoplasma gondii

This review summarises existing knowledge on the development and use of the polymerase chain reaction for the detection of DNA from Neospora and Toxoplasma. Several strategies which utilise the polymerase chain reaction for the diagnosis of toxoplasmosis in humans and livestock have been described and they principally target the B1 repetitive sequence, the P30 gene or ribosomal DNA. Experience has shown that the polymerase chain reaction has proven insufficiently robust to serve as a diagnostic test alone although when used in conjunction with other diagnostic techniques it does prove to be a useful aid. The marketing of a commercial polymerase chain reaction kit may well solve some of the inadequacies seen using ‘‘home made’’ polymerase chain reaction technology which are commonly used in diagnostic laboratories around the world. Recent progress on the development of polymerase chain reaction diagnostics for Neospora has been rapid and is discussed in detail.

Detection of Toxoplasma gondii tachyzoites and bradyzoites in blood, urine, and brains of infected mice.

Different techniques for identifying Toxoplasma gondii were compared. PCR was used to amplify part of the major surface antigen P30 gene of T. gondii. Amplified-DNA detection with the DNA enzyme immunoassay (PCR-DEIA) was more sensitive than ethidium bromide staining after agarose gel electrophoresis and as sensitive as nested PCR. PCR-DEIA, using common enzyme-linked immunosorbent assay (ELISA) methods, avoids agarose gel electrophoresis for the identification of amplified products. T. gondii can also be detected with equal sensitivity in infected fibroblasts, but only after at least 8 days of cell culture. PCR-DEIA is thus recommended because of its sensitivity and convenience for detecting early parasitemia in the surveillance of toxoplasmosis among pregnant women and immunocompromised hosts. The courses of infection in mice infected with two strains of T. gondii were compared. Tachyzoites of the virulent strain T. gondii RH, killing the host in 4 days, were identified in urine specimens and blood samples of mice 24 to 94 h after inoculation but not in brains, but no antibodies were detected. After intraperitoneal inoculation with cysts of the low-level virulence Beverley strain of T. gondii, parasites were identified in blood samples 4 days later and up to 17 days (but not in urine specimens) and in the brain from day 6 through day 525. By ELISA, high antibody titers were found from day 11 to day 525, with parasitemia preceding the appearance of antibodies. The usefulness of PCR-DEIA tests in conjunction with the search for circulating antibodies for the early diagnosis of toxoplasmosis in humans is discussed.

An overview of serological tests currently available for laboratory diagnosis of parasitic infections

Current methods and commercial test systems for the diagnosis of parasitic infections in both animals and humans are reviewed. Lists of test kits and their manufacturers are provided along with ordering information: the only commercially available test kits are for the diagnosis of toxoplasmosis in humans or animals and dirofilariasis (heartworm) in dogs. A partial list of diagnostic laboratories and the parasite tests they perform is also provided. Complete lists of diagnostic tests that could be obtained in the private sector are not available but would be useful. Two microfluorometric solid-phase assay systems are reviewed, and adaptations to custom assays for several kinds of parasites are briefly described. User problems in performing tests and interpreting results are stressed with emphasis placed on diagnosis of dirofilariasis in dogs. False-positive serology in dogs without heartworms and negative antibody reponses in micro-filariae-positive animals are discussed with respect to proper interpretation of results.