| Title: |
Multicenter evaluation of the Toxoplasma gondii Real-TM (Sacace) kit performance for the molecular diagnosis of toxoplasmosis |
| Authors: |
Guitard, Juliette; Brenier-Pinchart, Marie-Pierre; Varlet-Marie, Emmanuelle; Dalle, Frédéric; Rouges, Célia; Argy, Nicolas; Bonhomme, Julie; Capitaine, Agathe; Guégan, Hélène; Lavergne, Rose-Anne; Dardé, Marie-Laure; Pelloux, Hervé; Robert-Gangneux, Florence; Yera, Hélène; Sterkers, Yvon |
| Contributors: |
Mucoviscidose: physiopathologie et phénogénomique CRSA; Centre de Recherche Saint-Antoine (CRSA); Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU); CHU Saint-Antoine AP-HP; Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU); Centre National de Référence (CNR) Toxoplasmose/Toxoplasma Biological Resource Center (BRC) (CNR Toxoplasmose-Toxoplasma BRC); CHU Limoges; CHU de Grenoble-Alpes - Centre Hospitalier Universitaire CHU Grenoble (CHUGA); Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle (MIVEGEC); Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD Occitanie )-Université de Montpellier (UM); Laboratoire de parasitologie mycologie (CHU de Dijon); Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon); Service de parasitologie-mycologie CHU Cochin; Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin AP-HP; Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP); AP-HP - Hôpital Bichat - Claude Bernard Paris; CHU Caen Normandie – Centre Hospitalier Universitaire de Caen Normandie (CHU Caen Normandie); Normandie Université (NU); Service de Parasitologie-Mycologie Rennes; Université de Rennes (UR)-Centre Hospitalier Universitaire de Rennes CHU Rennes = Rennes University Hospital Pontchaillou; Cibles et Médicaments des Infections et de l'Immunité - UR 1155 (IICiMed); Nantes Université - UFR des Sciences Pharmaceutiques et Biologiques (Nantes Univ - UFR Pharmacie); Nantes Université - pôle Santé; Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ)-Nantes Université - pôle Santé; Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ); Service de Parasitologie Mycologie CHU Limoges; Institut de recherche en santé, environnement et travail (Irset); Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes (Biosit : Biologie - Santé - Innovation Technologique); Centre Hospitalier Universitaire de Rennes CHU Rennes = Rennes University Hospital Pontchaillou; Laboratoire de Parasitologie-Mycologie (CHU de Montpellier); Pôle Biologie-Pathologie CHRU Montpellier; Centre Hospitalier Régional Universitaire Montpellier (CHRU Montpellier)-Centre Hospitalier Régional Universitaire Montpellier (CHRU Montpellier); Centre Hospitalier Régional Universitaire Montpellier (CHRU Montpellier); Extramural funding was received from "Santé publique France" through the French National Reference Center for leishmaniasis. The funder had no role in study design, data collection and interpretation, or the decision to submit the work for publication. We are grateful to Sylvie Douzou, and France Joullié (Montpellier) for their technical help.We are grateful to the technical staff for their technical help (Sorbonne Université, APHP, Hôpital St Antoine,Laboratoire commun de génétique et biologie moléculaire). The authors would like to thank Valérie Martin, Mélanie Decara and Florent Saragosa (Grenoble, France) for their technical support. |
| Source: |
ISSN: 0095-1137 ; Journal of Clinical Microbiology ; https://hal.science/hal-04516294 ; Journal of Clinical Microbiology, 2024, 62 (4), pp.e0142823. ⟨10.1128/jcm.01428-23⟩. |
| Publisher Information: |
CCSD; American Society for Microbiology |
| Publication Year: |
2024 |
| Collection: |
Université de Nantes: HAL-UNIV-NANTES |
| Subject Terms: |
Toxoplasma gondii; comparaison methods; molecular diagnosis; [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology; [SDV.IB]Life Sciences [q-bio]/Bioengineering; [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases |
| Description: |
International audience ; ABSTRACT The molecular detection of Toxoplasma gondii DNA is a key tool for the diagnosis of disseminated and congenital toxoplasmosis. This multicentric study from the Molecular Biology Pole of the French National Reference Center for toxoplasmosis aimed to evaluate Toxoplasma gondii Real-TM PCR kit (Sacace). The study compared the analytical and clinical performances of this PCR assay with the reference PCRs used in proficient laboratories. PCR efficiencies varied from 90% to 112%; linearity zone extended over four log units (R 2 > 0.99) and limit of detection varied from 0.01 to ≤1 Tg/mL depending on the center. Determined on 173 cryopreserved DNAs from a large range of clinical specimens, clinical sensitivity was 100% [106/106; 95 confidence interval (CI): 96.5%–100%] and specificity was 100% (67/67; 95 CI: 94.6%–100%). The study revealed two potential limitations of the Sacace PCR assay: the first was the inconsistency of the internal control (IC) when added to the PCR mixture. This point was not found under routine conditions when the IC was added during the extraction step. The second is a lack of practicality, as the mixture is distributed over several vials, requiring numerous pipetting operations. Overall, this study provides useful information for the molecular diagnosis of toxoplasmosis; the analytical and clinical performances of the Sacace PCR kit were satisfactory, the kit having sensitivity and specificity similar to those of expert center methods and being able to detect low parasite loads, at levels where multiplicative analysis gives inconsistently positive results. Finally, the study recommends multiplicative analysis in particular for amniotic fluids, aqueous humor, and other single specimens. |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| Relation: |
info:eu-repo/semantics/altIdentifier/pmid/38470023; PUBMED: 38470023 |
| DOI: |
10.1128/jcm.01428-23 |
| Availability: |
https://hal.science/hal-04516294; https://hal.science/hal-04516294v1/document; https://hal.science/hal-04516294v1/file/SacaceGuitardetalPBM_vHAL240313.pdf; https://doi.org/10.1128/jcm.01428-23 |
| Rights: |
https://about.hal.science/hal-authorisation-v1/ ; info:eu-repo/semantics/OpenAccess |
| Accession Number: |
edsbas.1453E158 |
| Database: |
BASE |