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Identification of chromosomally integrated human herpesvirus 6 by droplet digital PCR.
- Source :
-
Clinical chemistry [Clin Chem] 2014 May; Vol. 60 (5), pp. 765-72. Date of Electronic Publication: 2014 Mar 04. - Publication Year :
- 2014
-
Abstract
- Background: Human herpesvirus 6 (HHV-6) latently infects a majority of adults. In about 1% of the population HHV-6 exists in a chromosomally integrated form (ciHHV-6) that resides in every somatic and germ cell and can be transmitted through the germ line. Patients with ciHHV-6 have been misdiagnosed and unnecessarily treated for active HHV-6 infection, sometimes with important side effects, based on results from quantitative molecular HHV-6 tests.<br />Methods: A droplet digital PCR (ddPCR) assay was developed to identify ciHHV-6 in cellular patient samples by precisely determining the ratio of HHV-6 to cellular DNA. We validated the assay on confirmed ciHHV-6 patient samples and a cell line derived from a ciHHV-6 patient, and we analyzed hematopoietic stem cell transplant patients suspected of having ciHHV-6. We additionally evaluated whether the assay could be applied to stored plasma samples from a study of clinical correlates of HHV-6.<br />Results: The ddPCR assay accurately identified ciHHV-6 in cellular samples (buffy coat, peripheral blood mononuclear cells), giving a ratio very close to 1 HHV-6/cell [mean (SD), 1.02 (0.03)] in fluorescence in situ hybridization-confirmed sample). In stored plasma samples, the assay performance was set by design to have 100% sensitivity, which resulted in 82% specificity for ciHHV-6.<br />Conclusions: The possibility of ciHHV-6 is often overlooked in patients with detectable HHV-6 viral loads by quantitative PCR. Our ddPCR test provides rapid and accurate laboratory identification of ciHHV-6 from easily obtained cellular samples. In addition, the assay provides excellent sensitivity and specificity using stored plasma samples, facilitating retrospective analysis of the clinical significance of ciHHV-6.
- Subjects :
- B-Lymphocytes virology
Cell Line
Hematopoietic Stem Cell Transplantation
Humans
In Situ Hybridization, Fluorescence
Reproducibility of Results
Roseolovirus Infections virology
Sensitivity and Specificity
Chromosomes, Human genetics
Chromosomes, Human virology
Herpesvirus 6, Human genetics
Herpesvirus 6, Human isolation & purification
Real-Time Polymerase Chain Reaction methods
Virus Integration genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1530-8561
- Volume :
- 60
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Clinical chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 24594780
- Full Text :
- https://doi.org/10.1373/clinchem.2013.217240