Torquetenovirus Viremia Quantification Using Real-Time PCR Developed on a Fully Automated, Random-Access Platform
Pietro Giorgio Spezia,
Fabrizio Carletti,
Federica Novazzi,
Eliana Specchiarello,
Angelo Genoni,
Francesca Drago Ferrante,
Claudia Minosse,
Giulia Matusali,
Nicasio Mancini,
Daniele Focosi,
Guido Antonelli,
Enrico Girardi,
Fabrizio Maggi
Affiliations
Pietro Giorgio Spezia
Laboratory of Virology, National Institute for Infectious Diseases Lazzaro Spallanzani—IRCCS, 00149 Rome, Italy
Fabrizio Carletti
Laboratory of Virology, National Institute for Infectious Diseases Lazzaro Spallanzani—IRCCS, 00149 Rome, Italy
Federica Novazzi
Laboratory of Medical Microbiology and Virology, Department of Medicine and Technological Innovation, Italy; Ospedale di Circolo e Fondazione Macchi, University of Insubria, 21100 Varese, Italy
Eliana Specchiarello
Laboratory of Virology, National Institute for Infectious Diseases Lazzaro Spallanzani—IRCCS, 00149 Rome, Italy
Angelo Genoni
Laboratory of Medical Microbiology and Virology, Department of Medicine and Technological Innovation, Italy; Ospedale di Circolo e Fondazione Macchi, University of Insubria, 21100 Varese, Italy
Francesca Drago Ferrante
Laboratory of Medical Microbiology and Virology, Department of Medicine and Technological Innovation, Italy; Ospedale di Circolo e Fondazione Macchi, University of Insubria, 21100 Varese, Italy
Claudia Minosse
Laboratory of Virology, National Institute for Infectious Diseases Lazzaro Spallanzani—IRCCS, 00149 Rome, Italy
Giulia Matusali
Laboratory of Virology, National Institute for Infectious Diseases Lazzaro Spallanzani—IRCCS, 00149 Rome, Italy
Nicasio Mancini
Laboratory of Medical Microbiology and Virology, Department of Medicine and Technological Innovation, Italy; Ospedale di Circolo e Fondazione Macchi, University of Insubria, 21100 Varese, Italy
Daniele Focosi
North-Western Tuscany Blood Bank, Pisa University Hospital, 56124 Pisa, Italy
Guido Antonelli
Laboratory of Microbiology and Virology, Department of Molecular Medicine, Sapienza University of Rome, 00185 Rome, Italy
Enrico Girardi
Scientific Direction, National Institute for Infectious Diseases Lazzaro Spallanzani—IRCCS, 00149 Rome, Italy
Fabrizio Maggi
Laboratory of Virology, National Institute for Infectious Diseases Lazzaro Spallanzani—IRCCS, 00149 Rome, Italy
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.
