Simultaneous Quantification of Epstein-Barr Virus, Cytomegalovirus, and Human Herpesvirus 6 DNA in Samples from Transplant Recipients by Multiplex Real-Time PCR Assay
Open Access
- 1 May 2007
- journal article
- research article
- Published by American Society for Microbiology in Journal of Clinical Microbiology
- Vol. 45 (5), 1426-1432
- https://doi.org/10.1128/jcm.01515-06
Abstract
We developed a multiplex real-time PCR assay using 6-carboxyfluorescein, 6-carboxy-4′,5′-dichloro-2′,7′-dimethoxyfluorescein, and carbocyanine 5-labeled probes to simultaneously quantify Epstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpesvirus 6 (HHV-6) DNA. When previously tested and stored DNA samples were examined, results of the multiplex real-time PCR assay were as sensitive and specific as those of a single real-time PCR assay. The multiplex assay was used to quantify the EBV, CMV, and HHV-6 DNA in 46 transplant recipients. A total of 303 whole-blood and plasma specimens were collected and analyzed. According to the results of the multiplex assay, the detection rates for viral DNA in whole blood and plasma were 23.8% and 5.9% for EBV, 11.2% and 5.3% for CMV, and 12.5% and 2.0% for HHV-6, respectively. All forms of viral DNA were detected more frequently in whole blood than in plasma. During the symptomatic period, EBV DNA was detected in all whole-blood specimens but not in all plasma specimens. Furthermore, the EBV DNA load in whole blood was higher during the symptomatic period than during the asymptomatic period, whereas the EBV DNA load in plasma was similar for both periods. These results demonstrate that whole blood is more suitable for the quantification of EBV DNA in transplant patients. However, a cutoff value with clinical relevance still needs to be determined.Keywords
This publication has 28 references indexed in Scilit:
- Herpesvirus Infections in Solid Organ Transplant Patients at High Risk of Primary Cytomegalovirus DiseaseThe Journal of Infectious Diseases, 2005
- High Fatality Rate of Epstein-Barr Virus-Associated Lymphoproliferative Disorder Occurring after Bone Marrow Transplantation with Rabbit Antithymocyte Globulin Conditioning RegimensJournal of Clinical Microbiology, 2005
- Severe Amnesia due to HHV-6 Encephalitis after Allogenic Stem Cell TransplantationEuropean Neurology, 2005
- Measurement of Epstein-Barr Virus DNA Loads in Whole Blood and Plasma by TaqMan PCR and in Peripheral Blood Lymphocytes by Competitive PCRJournal of Clinical Microbiology, 2003
- Real-Time PCR as a Versatile Tool for Investigating the Susceptibility of Human Herpesvirus 6 to Antiviral AgentsAntimicrobial Agents and Chemotherapy, 2003
- Automated Extraction and Quantification of Human Cytomegalovirus DNA in Whole Blood by Real-Time PCR AssayJournal of Clinical Microbiology, 2003
- Herpesvirus Infections in Organ Transplant RecipientsClinical and Vaccine Immunology, 2003
- Cytomegalovirus (CMV) DNA Load in Plasma for the Diagnosis of CMV Disease before Engraftment in Hematopoietic Stem‐Cell Transplant RecipientsThe Journal of Infectious Diseases, 2001
- Monitoring four herpesviruses in unrelated cord blood transplantationBone Marrow Transplantation, 2000
- Herpesviruses.Journal of Clinical Pathology, 1979