Detection of Toxoplasma gondii from amniotic fluid, a comparison of four different molecular biological methods

Nagy, Béla; Bán, Zoltán; Beke, Artúr; Nagy, Gyula Richárd; Lázár, Levente; Papp, Csaba; Tóth-Pál, E.; Papp, Zoltán

doi:10.1016/j.cca.2005.12.023

Cited by 27 publications

(22 citation statements)

References 25 publications

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“…This real-time PCR format was used because of its popularity in the analysis of clinical specimens (5, 7, 10-13, 20, 24, 29, 37, 38, 41, 43, 45), the time-saving nature, and the potential for quantitation (4). FRET hybridization probes were used instead of SYBR green in the detection step of the PCR assay to increase sensitivity and specificity (19,43).…”

Section: Discussionmentioning

confidence: 99%

Detection of Toxoplasma gondii Oocysts in Water Sample Concentrates by Real-Time PCR

Yang

Lindquist

Cama

et al. 2009

Appl Environ Microbiol

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PCR techniques in combination with conventional parasite concentration procedures have potential for the sensitive and specific detection of Toxoplasma gondii oocysts in water. Three real-time PCR assays based on the B1 gene and a 529-bp repetitive element were analyzed for the detection of T. gondii tachyzoites and oocysts. Lower sensitivity and specificity were obtained with the B1 gene-based PCR than with the 529-bp repeat-based PCR. New procedures for the real-time PCR detection of T. gondii oocysts in concentrates of surface water were developed and tested in conjunction with a method for the direct extraction of inhibitor-free DNA from water. This technique detected as few as one oocyst seeded to 0.5 ml of packed pellets from water samples concentrated by Envirocheck filters. Thus, this real-time PCR may provide a detection method alternative to the traditional mouse assay and microscopy.

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Section: Discussionmentioning

confidence: 99%

Detection of Toxoplasma gondii Oocysts in Water Sample Concentrates by Real-Time PCR

Yang

Lindquist

Cama

et al. 2009

Appl Environ Microbiol

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“…A similar discrepancy between nested and real-time PCR results has been reported previously. [30][31][32] In primary toxoplasmosis or reactivation among liver recipients, total antibody levels have been reported to slightly increase or even decrease because of leukoneutropenia due to intense immunosuppressive treatment. 2,3,57,10,12,19,29,33 In the present study, according to the results of the serological assays, IgG and IgM levels increased only in PCR-positive patient 4, IgG levels decreased in PCR-positive patient 17, and antibody levels remained unchanged in PCR-positive patient 13 (Table 2).…”

Section: Discussionmentioning

confidence: 99%

Incidence and diagnosis of active toxoplasma infection among liver transplant recipients in Western Turkey

et al. 2008

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Toxoplasmosis is a serious and potentially life-threatening disease in liver transplant recipients while they are immunosuppressed. We report the clinical and laboratory findings related to active toxoplasma infection associated with 40 immunosuppressed liver transplant procedures that took place over a 12-month period at a major transplant unit in Izmir, Turkey. Twenty-seven (67.5%) of the 40 transplant recipients were found to be seropositive for toxoplasma infection and therefore at risk of reactivated infection. From the serological status of the donors, which was ascertained in 38 of 40 cases, we identified 3 (7.9%) of 38 transplants to be from a seropositive donor to a seronegative recipient. In 10 (26.3%) of 38 transplants, both the donor and recipient were seronegative, and this excluded toxoplasma as a risk. A comparison of real-time polymerase chain reaction (PCR) and nested PCR was undertaken in combination with a range of serological assays (the Sabin-Feldman dye test, enzyme immunoassay immunoglobulin M, and immunosorbent agglutination assay immunoglobulin M). Ethylene diamine tetraacetic acid blood samples from 3 of the 30 recipients at risk from toxoplasma were found positive by PCR, but only 1 of these was found positive in both assays. Among the 3 PCR-positive patients, immunoglobulin M and immunoglobulin G antibody levels increased in only 1 patient. Correlations between symptoms, laboratory findings, and clinical management (use of anti-toxoplasma therapy) are presented. Our findings suggest that toxoplasma presents a significant risk to our liver transplant population and that PCR is a helpful addition in identifying active infections and hence in informing clinical management decisions. Liver Transpl 14:1526Transpl 14: -1532Transpl 14: , 2008

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“…Thus, for Toxoplasma gondii, another protozoan parasite, qrtPCR assays using exactly the same (B1 gene) target and different primers have been reported with analytical sensitivities varying by a 200-fold factor, i.e., from 0.05 to 10 genome equivalents per reaction tube (6,7,11,13,14,19,25,33,36). Still, such variations are due not only to the choice of the primers but also, at least as importantly, to the optimization of the method: indeed, identical primers may yield widely different performances in different laboratories (P. Bastien and the Molecular Biology Group of the French National Reference Center for Toxoplasmosis, unpublished data).…”

mentioning

confidence: 99%

Quantitative Real-Time PCR Is Not More Sensitive than “Conventional” PCR

2008

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Detection of Toxoplasma gondii from amniotic fluid, a comparison of four different molecular biological methods

Cited by 27 publications

References 25 publications

Detection of Toxoplasma gondii Oocysts in Water Sample Concentrates by Real-Time PCR

Detection of Toxoplasma gondii Oocysts in Water Sample Concentrates by Real-Time PCR

Incidence and diagnosis of active toxoplasma infection among liver transplant recipients in Western Turkey

Quantitative Real-Time PCR Is Not More Sensitive than “Conventional” PCR

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