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Journal of Clinical Microbiology, August 2004, p. 3681-3685, Vol. 42, No. 8
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.8.3681-3685.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Use of Fluorescence Resonance Energy Transfer Hybridization Probes To Evaluate Quantitative Real-Time PCR for Diagnosis of Ocular Toxoplasmosis

Audrey Simon,1 Pierre Labalette,2 Isabelle Ordinaire,1 Emilie Fréalle,1,3 Eduardo Dei-Cas,1,3 Daniel Camus,1,3 and Laurence Delhaes1,3*

Parasitology-Mycology Department,1 Ophthalmology Department, Lille 2 University Hospital Center,2 EA 3609, Lille Pasteur Institute, Lille, France3

Received 20 February 2004/ Returned for modification 4 April 2004/ Accepted 30 April 2004

Toxoplasma gondii infection is an important cause of chorioretinitis in Europe and the United States. Ophthalmological examination and a good clinical response to adequate therapy mainly support ocular toxoplasmosis diagnosis. However, clinical diagnostic may be difficult in some atypical cases. In these cases, laboratory confirmation, based on detection of local specific antibodies and parasite DNA by conventional PCR, is therefore important to confirm the disease etiology. More recently, real-time PCR has been developed to improve prenatal congenital toxoplasmosis diagnosis. We therefore examined the diagnostic value of quantitative real-time PCR for the detection of T. gondii in aqueous humor samples, associated with quantification of human ß-globin to control sample quantitative quality, by using a double fluorescence resonance energy transfer hybridization probes system with a double fluorescence reading. Of the 23 the clinically toxoplasmosis suspect patients, 22 showed serological evidence of exposure to Toxoplasma; one had a serological profile indicative of active infection. The analysis of paired aqueous humor and serum samples revealed an intraocular antibody production in 9 of 23 cases (39.1%). The quantitative real-time PCR revealed positive and high parasite numbers and high Toxoplasma/human genome ratios in three cases. Furthermore, PCR was the only positive confirmatory test in two cases (11.1%). None of the patients included in the control group (n = 7) had evidence of either local specific antibody production or T. gondii DNA detection, suggesting a good relative assay specificity. On the whole, quantitative real-time PCR appears to be useful for diagnosing atypical ocular toxoplasmosis presentations.

* Corresponding author. Mailing address: Service de Parasitologie-Mycologie, Faculté de Médecine, Hôpital Huriez, 1 Place de Verdun, 59045 Lille Cedex 2, France. Phone: 33-3-20-44-55-77. Fax: 33-3-20-44-42-64. E-mail: l-delhaes{at}chru-lille.fr.

Journal of Clinical Microbiology, August 2004, p. 3681-3685, Vol. 42, No. 8
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.8.3681-3685.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Fekkar, A., Bodaghi, B., Touafek, F., Le Hoang, P., Mazier, D., Paris, L. (2008). Comparison of Immunoblotting, Calculation of the Goldmann-Witmer Coefficient, and Real-Time PCR Using Aqueous Humor Samples for Diagnosis of Ocular Toxoplasmosis. J. Clin. Microbiol. 46: 1965-1967 [Abstract] [Full Text]  
  • Cassaing, S., Bessieres, M. H., Berry, A., Berrebi, A., Fabre, R., Magnaval, J. F. (2006). Comparison between Two Amplification Sets for Molecular Diagnosis of Toxoplasmosis by Real-Time PCR.. J. Clin. Microbiol. 44: 720-724 [Abstract] [Full Text]  


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