PCR-Based Detection of Toxoplasma gondii DNA in Blood and Ocular Samples for Diagnosis of Ocular Toxoplasmosis

Bourdin, C.; Busse, Andreas; Kouamou, E.; Touafek, Fériel; Bodaghi, Bahram; Hoang, P. Le; Mazier, Dominique; Paris, Luc; Fekkar, Arnaud

doi:10.1128/jcm.01793-14

Cited by 58 publications

(23 citation statements)

References 24 publications

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“…This patient shows a different profile since he presented serological evidence of acute infection but the cPCR was negative during the follow up. Maybe this is a case of atypical strain infection not detected by B1 gene as previously described and the cPCR should be done using other set of primer as being reported in other regions of the world [ 3 , 26 , 44 , 47 , 63 – 67 ], but not in South American studies [ 30 , 45 , 48 – 50 , 68 – 72 ].…”

Section: Discussionmentioning

confidence: 92%

A Brazilian report using serological and molecular diagnosis to monitoring acute ocular toxoplasmosis

et al. 2015

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BackgroundToxoplasmosis was recently included as a neglected disease by the Center for Disease Control. Ocular toxoplasmosis is one clinical presentation of congenital or acquired infection. The laboratory diagnosis is being used worldwide to support the clinical diagnosis and imaging. The aim of this study was to evaluate the use of serology and molecular methods to monitor acute OT in immunocompetent patients during treatment.MethodsFive immunocompetent patients were clinically diagnosed with acute OT. The clinical evaluation was performed by ophthalmologic examination using the Early Treatment Diabetic Retinopathy Study, best-corrected visual acuity, slit lamp biomicroscopy, fundoscopic examination with indirect binocular ophthalmoscopy color fundus photography, fluorescein angiography and spectral optical coherence tomography (OCT). Serology were performed by ELISA (IgA, IgM, IgG) and confirmed by ELFA (IgG, IgM). Molecular diagnoses were performed in peripheral blood by cPCR using the Toxoplasma gondiiB1 gene as the marker. Follow-up exams were performed on day +15 and day +45.ResultsOnly five non-immunocompromised male patients completed the follow up and their data were used for analysis. The mean age was 41.2 ± 11.3 years (median: 35; range 31–54 years). All of them were positive for IgG antibodies but with different profiles for IgM and IgA, as well as PCR. For all patients the OCT exam showed active lesions with the inner retinal layers being abnormally hyper-reflective with full-thickness disorganization of the retinal reflective layers, which assumed a blurred reflective appearance and the retina was thickened.ConclusionsThe presence of IgA and IgM confirmed the acute infection and thus was in agreement with the clinical evaluation. Our results show the adopted treatment modified the serological profile of IgM antibodies and the PCR results, but not the IgG and IgA antibodies and that imaging is a good tool to follow-up patients.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-015-1650-6) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 92%

A Brazilian report using serological and molecular diagnosis to monitoring acute ocular toxoplasmosis

et al. 2015

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“…For example, the sensitivity of PCR in blood samples is very low for diagnosing ocular toxoplasmosis in immunocompetent patients from France and T . gondii DNA is detectable only in ocular fluid samples [ 40 , 41 ]. In contrast, T .…”

Section: Discussionmentioning

confidence: 99%

Performance Testing of PCR Assay in Blood Samples for the Diagnosis of Toxoplasmic Encephalitis in AIDS Patients from the French Departments of America and Genetic Diversity of Toxoplasma gondii: A Prospective and Multicentric Study

Ajzenberg

Lamaury

Demar³

et al. 2016

PLoS Negl Trop Dis

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BackgroundToxoplasmic encephalitis in patients with AIDS is a life-threatening disease mostly due to reactivation of Toxoplasma gondii cysts in the brain. The main objective of this study was to evaluate the performance of real-time PCR assay in peripheral blood samples for the diagnosis of toxoplasmic encephalitis in AIDS patients in the French West Indies and Guiana.Methodology/Principal FindingsAdult patients with HIV and suspicion of toxoplasmic encephalitis with start of specific antitoxoplasmic therapy were included in this study during 40 months. The real-time PCR assay targeting the 529 bp repeat region of T. gondii was performed in two different centers for all blood samples. A Neighbor-Joining tree was reconstructed from microsatellite data to examine the relationships between strains from human cases of toxoplasmosis in South America and the Caribbean. A total of 44 cases were validated by a committee of experts, including 36 cases with toxoplasmic encephalitis. The specificity of the PCR assay in blood samples was 100% but the sensitivity was only 25% with moderate agreement between the two centers. Altered level of consciousness and being born in the French West Indies and Guiana were the only two variables that were associated with significantly decreased risk of false negative results with the PCR assay.Conclusion/SignificanceOur results showed that PCR sensitivity in blood samples increased with severity of toxoplasmic encephalitis in AIDS patients. Geographic origin of patients was likely to influence PCR sensitivity but there was little evidence that it was caused by differences in T. gondii strains.Trial RegistrationClinicalTrials.gov NCT00803621

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“…PCR has been found to detect T. gondii DNA more effectively in aqueous humor (25%) than in peripheral blood (5%) samples (22) . Recently, a study demonstrated that the sensitivity for blood was poor (4.1%) compared with that for ocular samples (35.9%) (23) . These findings corroborate our results, which showed that the ability of PCR to detect T. gondii DNA was more sensitive in aqueous humor (37.21%) than in peripheral blood samples (2.33%).…”

Section: Discussionmentioning

confidence: 99%

Detection ofToxoplasma gondiiDNA in peripheral blood and aqueous humor of patients with Toxoplasmic active focal necrotizing retinochoroiditis using real-time PCR

Santos¹,

Nascimento²,

Muccioli³

et al. 2015

Arquivos Brasileiros De Oftalmologia

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Purpose: To evaluate the ability of real-time quantitative PCR (qPCR) for detecting Toxoplasma gondii DNA in the peripheral blood and aqueous humor of patients with toxoplasmic active focal necrotizing retinochoroiditis. Methods: Fifty-five patients with infectious uveitis seen from 2009 to 2013 at the Department of Ophthalmology and Visual Sciences of the Federal University of São Paulo were enrolled in this study. Forty-three patients had toxoplasmic active focal necrotizing retinochoroiditis, and the remaining 12 had non-toxoplasmic infectious uveitis and served as controls. qPCR analysis for T. gondii DNA was performed on the patients' peripheral blood and aqueous humor samples. Results:The qPCR was positive for T. gondii DNA in 37.21% (16/43) of the aqueous humor samples and 2.33% (1/43) of the peripheral blood samples; further, 16.27% (7/43) of the patients had positive results in both their blood and aqueous humor samples. Conclusion: qPCR was able to detect T. gondii DNA in patients with toxoplasmic active focal necrotizing retinochoroiditis in the blood as well as the aqueous humor and can help with the diagnosis of the disease.

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PCR-Based Detection of Toxoplasma gondii DNA in Blood and Ocular Samples for Diagnosis of Ocular Toxoplasmosis

Cited by 58 publications

References 24 publications

A Brazilian report using serological and molecular diagnosis to monitoring acute ocular toxoplasmosis

A Brazilian report using serological and molecular diagnosis to monitoring acute ocular toxoplasmosis

Performance Testing of PCR Assay in Blood Samples for the Diagnosis of Toxoplasmic Encephalitis in AIDS Patients from the French Departments of America and Genetic Diversity of Toxoplasma gondii: A Prospective and Multicentric Study

Detection ofToxoplasma gondiiDNA in peripheral blood and aqueous humor of patients with Toxoplasmic active focal necrotizing retinochoroiditis using real-time PCR

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