Molecular Diagnosis of Toxoplasmosis : Multicenter Evaluation of the Toxoplasma RealCycler Universal PCR Assay on 168 Characterized Human Samples - Université de Rennes Accéder directement au contenu
Article Dans Une Revue Journal of Molecular Diagnostics Année : 2022

Molecular Diagnosis of Toxoplasmosis : Multicenter Evaluation of the Toxoplasma RealCycler Universal PCR Assay on 168 Characterized Human Samples

Résumé

Real-time PCR plays a crucial role in the diagnosis of toxoplasmosis. In this multicenter study, the Toxoplasma RealCycler Universal assay was assessed for the diagnosis of toxoplasmosis by eight reference laboratories. DNAs from diverse clinical samples were included: 141 characterized samples from patients with different clinical forms of proven toxoplasmosis and 27 from patients without toxoplasmosis were tested in duplicate with the commercial assay. Final diagnosis was affirmed by each center by analysis of clinical settings and biological follow-up. Calibrated Toxoplasma gondii standards and 11 external quality control samples were also included. Discrepant results observed after the first run of commercial PCR were controlled by both reference and commercial PCR assays. Using the commercial assay, the detection threshold varied from 0.01 to 1 tachyzoites/mL, depending on the center. The relationship between crossing point and DNA concentration was linear over 4 log units (r2 > 0.99), and PCR efficiencies were satisfactory (89% to 104%). The results of the 11 external quality control samples were concordant after one retesting, but those for 3 clinical samples remained discrepant. Sensitivity and specificity were calculated at 97.8% (97.8% to 100%) and 100% (87.2% to 100%), respectively. Provided that PCRs are performed at least in duplicate to detect low parasitic loads, Toxoplasma RealCycler Universal PCR showed suitable performances to diagnose the different forms of toxoplasmosis.
Fichier principal
Vignette du fichier
Progenie-Brenier Pinchart et al -JMDi.pdf (751.74 Ko) Télécharger le fichier
Origine : Fichiers produits par l'(les) auteur(s)

Dates et versions

hal-03688137 , version 1 (03-06-2022)

Identifiants

Citer

Marie-Pierre Brenier-Pinchart, Denis Filisetti, Sophie Cassaing, Emmanuelle Varlet-Marie, Florence Robert-Gangneux, et al.. Molecular Diagnosis of Toxoplasmosis : Multicenter Evaluation of the Toxoplasma RealCycler Universal PCR Assay on 168 Characterized Human Samples. Journal of Molecular Diagnostics, 2022, 24 (6), pp.687-696. ⟨10.1016/j.jmoldx.2022.03.009⟩. ⟨hal-03688137⟩
105 Consultations
77 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More