Simultaneous Detection of Beta and Gamma Human Herpesviruses by Multiplex qPCR Reveals Simple Infection and Coinfection Episodes Increasing Risk for Graft Rejection in Solid Organ Transplantation

• Published in: Viruses 2018-12, Vol.10 (12), p.730
• Main Authors: Sánchez-Ponce, Yessica, Varela-Fascinetto, Gustavo, Romo-Vázquez, José Carlos, López-Martínez, Briceida, Sánchez-Huerta, José Luis, Parra-Ortega, Israel, Fuentes-Pananá, Ezequiel M, Morales-Sánchez, Abigail
• Format: Article
• Language: English
• Subjects: Adolescent; Child; coinfection; Coinfection - diagnosis; Coinfection - virology; Cytomegalovirus; DNA Primers - genetics; DNA, Viral - blood; Enzymes; Epstein-Barr virus; Epstein-Barr Virus Infections - diagnosis; Epstein-Barr Virus Infections - virology; Female; Genes; Genomes; Graft rejection; Graft Rejection - etiology; Graft Rejection - virology; Herpesvirus 4, Human - genetics; Herpesvirus 6, Human - genetics; Herpesvirus 7, Human - genetics; human herpesviruses; Humans; Immunosurveillance; Infections; Kidney transplantation; Laboratories; Leukocytes; Liver transplantation; Lymphoma; Male; Microbiomes; Multiplex Polymerase Chain Reaction; Organ Transplantation - adverse effects; Pediatrics; Plasmids; Prospective Studies; Reproducibility; Roseolovirus Infections - diagnosis; Roseolovirus Infections - virology; Sarcoma; Sensitivity and Specificity; Software; transplantation; Transplants & implants; Viral Load
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v10120730

Herpesviruses are common components of the human microbiome that become clinically relevant when a competent immunosurveillance is compromised, such as in transplantation. Members of the beta and gamma subfamilies are associated with a wide diversity of pathologies, including end-organ disease and cancer. In this study, we developed a multiplex qPCR technique with high specificity, sensitivity, efficiency and predictability that allowed the simultaneous detection and quantification of beta and gamma human herpesviruses. The technique was tested in a cohort of 34 kidney- or liver-transplanted pediatric patients followed up for up to 12 months post-transplant. Viral load was determined in 495 leukocyte-plasma paired samples collected bi-weekly or monthly. Human herpesvirus (HHV) 7 was the herpesvirus most frequently found in positive samples (39%), followed by Epstein-Barr virus (EBV) (20%). Also, EBV and HHV7 were present in the majority of coinfection episodes (62%). The share of positive samples exclusively detected either in leukocytes or plasma was 85%, suggesting that these herpesviruses tended to take a latent or lytic path in an exclusive manner. Infection by human cytomegalovirus (HCMV) and HHV6, as well as coinfection by EBV/HHV7 and EBV/HHV6/HHV7, were associated with graft rejection (RR = 40.33 ( = 0.0013), 5.60 ( = 0.03), 5.60 ( = 0.03) and 17.64 ( = 0.0003), respectively). The routine monitoring of beta and gamma herpesviruses should be mandatory in transplant centers to implement preventive strategies.