1. Torquetenovirus Viremia Quantification Using Real-Time PCR Developed on a Fully Automated, Random-Access Platform.
- Author
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Spezia PG, Carletti F, Novazzi F, Specchiarello E, Genoni A, Drago Ferrante F, Minosse C, Matusali G, Mancini N, Focosi D, Antonelli G, Girardi E, and Maggi F
- Subjects
- Humans, Sensitivity and Specificity, Torque teno virus genetics, Torque teno virus isolation & purification, DNA, Viral genetics, DNA, Viral blood, Limit of Detection, Reproducibility of Results, Automation, Laboratory methods, Real-Time Polymerase Chain Reaction methods, Viremia diagnosis, Viremia virology, Viral Load methods, DNA Virus Infections diagnosis, DNA Virus Infections virology
- Abstract
Quantification of Torquetenovirus (TTV) viremia is becoming important for evaluating the status of the immune system in solid organ transplant recipients, monitoring the appearance of post-transplant complications, and controlling the efficacy of maintenance immunosuppressive therapy. Thus, diagnostic approaches able to scale up TTV quantification are needed. Here, we report on the development and validation of a real-time PCR assay for TTV quantification on the Hologic Panther Fusion
® System by utilizing its open-access channel. The manual real-time PCR previously developed in our laboratories was optimized to detect TTV DNA on the Hologic Panther Fusion® System. The assay was validated using clinical samples. The automated TTV assay has a limit of detection of 1.6 log copies per ml of serum. Using 112 samples previously tested via manual real-time PCR, the concordance in TTV detection was 93% between the assays. When the TTV levels were compared, the overall agreement between the methods, as assessed using Passing-Bablok linear regression and Bland-Altman analyses, was excellent. In summary, we validated a highly sensitive and accurate method for the diagnostic use of TTV quantification on a fully automated Hologic Panther Fusion® System. This will greatly improve the turnaround time for TTV testing and better support the laboratory diagnosis of this new viral biomarker.- Published
- 2024
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