Contributions of Immunoblotting, Real-Time PCR, and the Goldmann-Witmer Coefficient to Diagnosis of Atypical Toxoplasmic Retinochoroiditis

• Published in: Journal of Clinical Microbiology Vol. 47; no. 7; pp. 2131 - 2135
• Main Authors: Talabani, H, Asseraf, M, Yera, H, Delair, E, Ancelle, T, Thulliez, P, Brézin, A.P, Dupouy-Camet, J
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01-07-2009; American Society for Microbiology (ASM)
• Subjects: Adolescent; Adult; Aged; Aged, 80 and over; Animals; Biological and medical sciences; Choroiditis - diagnosis; Female; Fundamental and applied biological sciences. Psychology; Human protozoal diseases; Humans; Immunoblotting - methods; Infectious diseases; Male; Medical sciences; Microbiology; Middle Aged; Parasitic diseases; Parasitology; Polymerase Chain Reaction - methods; Protozoal diseases; Retinitis - diagnosis; Sensitivity and Specificity; Toxoplasma - immunology; Toxoplasma - isolation & purification; Toxoplasma gondii; Toxoplasmosis; Toxoplasmosis - complications; Toxoplasmosis, Ocular - diagnosis; Young Adult; Protozoal disease; Retinopathy; Microbiology; Uvea disease; Parasitosis; Chorioretinitis; Real time; Infection; Polymerase chain reaction; Eye disease; Toxoplasmosis; Retinochoroiditis; Immunoblotting assay; Atypical; Diagnosis
• ISSN: 0095-1137; 1098-660X
• DOI: 10.1128/JCM.00128-09

Ocular toxoplasmosis is a major cause of posterior uveitis worldwide. The diagnosis is based mainly on ophthalmological examination. Biological diagnosis is necessary in atypical cases, and this requires aqueous humor sampling by anterior chamber paracentesis. We evaluated real-time PCR targeting the Toxoplasma gondii 529-bp repeat element, the Goldmann-Witmer coefficient (GWC), and immunoblotting for the diagnosis of toxoplasmic retinochoroiditis in 54 patients with atypical uveitis. The results of these biological tests, applied to paired aqueous humor-serum samples, were compared to the clinical findings. Combining either PCR or the GWC with immunoblotting increased the sensitivity to 73% or 70%, respectively. Together, PCR and the GWC had 80% sensitivity. If feasible, sensitivity can be increased by combining the three methods (85% sensitivity). The interval between symptom onset and anterior chamber paracentesis strongly influenced the detection of specific intraocular antibody synthesis. The sensitivity of the GWC increased from 45% to 56% when sampling was performed 10 days after symptom onset, and that of immunoblotting increased from 53% to 72% when puncture was performed 30 days after symptom onset. PCR analysis of aqueous humor samples detected toxoplasmic DNA in 55% of patients. In contrast to the results of immunoblotting and the GWC, the results of PCR were not influenced by the interval between symptom onset and paracentesis. PCR was more informative than the GWC and immunoblotting for immunocompromised patients. Acute necrotizing retinal lesions were significantly larger in PCR-positive patients, with a mean of 3.5 optic disc diameters, than in PCR-negative patients, with a mean of 1.5 optic disc diameters.