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Virus Detection and Semiquantitation in Explanted Heart Tissues of Idiopathic Dilated Cardiomyopathy Adult Patients by Use of PCR Coupled with Mass Spectrometry Analysis
Authors:Yohan Nguyen  Fanny Renois  Nicolas Leveque  Delphine Giusti  Marcus Picard-Maureau  Patrick Bruneval  Paul Fornes  Laurent Andreoletti
Affiliation:Clinical and Molecular Virology Unit, University Hospital, and EA-4684 Cardiovir SFR-CAP Sante, Faculty of Medicine, Reims, Francea;Abbott GmbH & Co. KG, Wiesbaden, Germanyb;Pathology Department, HEGP University Hospital and Paris VI University, Paris, Francec;Pathology Department, University Hospital, and EA-4684 Cardiovir SFR-CAP Sante, Faculty of Medicine, Reims, Franced
Abstract:Viral detection in heart tissues has become a central issue for the diagnosis and exploration of the pathogenesis of idiopathic dilated cardiomyopathy (IDCM). In the present study, common cardiotropic viruses in 67 explanted heart samples of 31 IDCM adult patients were detected and semiquantified by using for the first time a new technology based on PCR assay coupled to electrospray ionization-time of flight mass spectrometry analysis (PCR-MS), with comparison to reference quantitative real-time PCR (RT-qPCR) assay. PCR-MS identified single or mixed enterovirus (EV) and parvovirus B19 (PVB19) infections in 27 (40.2%) of 67 samples, corresponding to 15 (48.3%) of the 31 patients, whereas RT-qPCR identified viral infections in 26 (38.8%) samples, corresponding to 16 (51.6%) of the patients. The PCR-MS results correlated well with EV and PVB19 detection by RT-qPCR (kappa = 0.85 95% confidence interval {CI}, 0.72 to 1.00] and kappa = 0.82 95% CI, 0.66 to 0.99], respectively). The levels of EV RNA (median, 550 range, 178 to 3,200] copies/μg of total extracted nucleic acids) and of PVB19 DNA (median, 486 range, 80 to 1,157] copies/μg of total extracted nucleic acids) were measured using PCR-MS and correlated with those obtained by RT-qPCR (r2 = 0.57, P = 0.002 and r2 = 0.64, P < 0.001 for EV and PVB19, respectively). No viruses other than EV and PVB19 strains were detected using the new PCR-MS technology, which is capable of simultaneously identifying 84 known human viruses in one assay. In conclusion, we identified single or mixed EV and PVB19 cardiac infections as potential causes of IDCM. The PCR-MS analysis appeared to be a valuable tool to rapidly detect and semiquantify common viruses in cardiac tissues and may be of major interest to better understand the role of viruses in unexplained cardiomyopathies.
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