Dorian Kulifaj1, Bénédicte Durgueil-Lariviere2, Faustine Meynier3, Eliza Munteanu4, Nicolas Pichon5, Manon Dubé6, Martine Joannes6, Marie Essig2, Sébastien Hantz4, Côme Barranger7, Sophie Alain8. 1. UMR INSERM 1092, Université de Limoges, National Reference Center For Herpesviruses, Bacteriology-Virology-Hygiene Department, CHU Limoges, 2 ave ML King, 87000, Limoges, France; bioMérieux, 138 rue Louis Pasteur, Parc technologique Delta Sud, 09340, Verniolle, France. 2. Nephrology and Transplantation Department, CHU Limoges, 2 ave ML King, 87000, Limoges, France. 3. bioMérieux, Centre Christophe Mérieux, 5 rue des Berges, 38024, Grenoble cedex 01, Grenoble, France. 4. UMR INSERM 1092, Université de Limoges, National Reference Center For Herpesviruses, Bacteriology-Virology-Hygiene Department, CHU Limoges, 2 ave ML King, 87000, Limoges, France. 5. Intensive Care Unit Department, CHU Limoges, 2 ave ML King, 87000, Limoges, France. 6. bioMérieux, 138 rue Louis Pasteur, Parc technologique Delta Sud, 09340, Verniolle, France. 7. bioMérieux, 138 rue Louis Pasteur, Parc technologique Delta Sud, 09340, Verniolle, France. Electronic address: come.barranger@biomerieux.com. 8. UMR INSERM 1092, Université de Limoges, National Reference Center For Herpesviruses, Bacteriology-Virology-Hygiene Department, CHU Limoges, 2 ave ML King, 87000, Limoges, France. Electronic address: sophie.alain@unilim.fr.
Abstract
BACKGROUND: Torque teno viruses (TTV) are small DNA viruses whose replication is closely linked to immune status. A growing number of publications underlined the potential of TTV viral load as an indicator of immunosuppression. OBJECTIVES: To demonstrate the analytical performance of the first standardized RUO (Research Use Only) assay to detect and quantify human TTV DNA in whole blood and plasma. STUDY DESIGN: We established analytical performances for TTV load measurement in various populations. The TTV kinetics were followed in kidney recipients. TTV viral load was analyzed on whole blood samples from 42 kidney recipients follow-up, 53 kidney deceased donors and 31 healthy volunteers. RESULTS: The qPCR TTV assay detects the most prevalent human TTV genotypes and does not cross react with other viruses. Limit of detection was 2.2 log10 copies/mL in whole blood and plasma, linearity and precision were demonstrated over the range 1.61 to 10.61 log10 copies/mL in whole blood. Prevalence of TTV DNA in blood differed significantly among groups: 45% in healthy volunteers, 74% in donors and 83% in kidney recipients. In kidney recipients, early TTV kinetics were comparable to those previously observed with in-house assays in other transplant settings: viral load increased from an average of 4.3 log10 to 7.9 log10 copies/mL within the first 75 days post transplantation. CONCLUSION: This TTV assay showed high analytical sensitivity, specificity, linearity and precision. It is a useful standardized tool to further evaluate TTV load as a biomarker of immune status that could improve individual treatment strategy.
BACKGROUND: Torque teno viruses (TTV) are small DNA viruses whose replication is closely linked to immune status. A growing number of publications underlined the potential of TTV viral load as an indicator of immunosuppression. OBJECTIVES: To demonstrate the analytical performance of the first standardized RUO (Research Use Only) assay to detect and quantify human TTV DNA in whole blood and plasma. STUDY DESIGN: We established analytical performances for TTV load measurement in various populations. The TTV kinetics were followed in kidney recipients. TTV viral load was analyzed on whole blood samples from 42 kidney recipients follow-up, 53 kidney deceased donors and 31 healthy volunteers. RESULTS: The qPCR TTV assay detects the most prevalent human TTV genotypes and does not cross react with other viruses. Limit of detection was 2.2 log10 copies/mL in whole blood and plasma, linearity and precision were demonstrated over the range 1.61 to 10.61 log10 copies/mL in whole blood. Prevalence of TTV DNA in blood differed significantly among groups: 45% in healthy volunteers, 74% in donors and 83% in kidney recipients. In kidney recipients, early TTV kinetics were comparable to those previously observed with in-house assays in other transplant settings: viral load increased from an average of 4.3 log10 to 7.9 log10 copies/mL within the first 75 days post transplantation. CONCLUSION: This TTV assay showed high analytical sensitivity, specificity, linearity and precision. It is a useful standardized tool to further evaluate TTV load as a biomarker of immune status that could improve individual treatment strategy.
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