Identification of Chromosomally Integrated Human Herpesvirus 6 by Droplet Digital PCR

被引：90

作者：

Sedlak, Ruth Hall ^{[1
]}

Cook, Linda ^{[1
]}

Huang, Meei-Li ^{[1
]}

Magaret, Amalia ^{[1
,4
]}

Zerr, Danielle M. ^{[2
,3
]}

Boeckh, Michael ^{[2
,4
]}

Jerome, Keith R. ^{[1
,4
]}

机构：

[1] Univ Washington, Med Ctr, Dept Lab Med, Seattle, WA 98195 USA

[2] Univ Washington, Med Ctr, Dept Med, Seattle, WA 98195 USA

[3] Seattle Childrens Hosp, Dept Pediat, Seattle, WA USA

[4] Fred Hutchinson Canc Res Ctr, Program Infect Dis, Vaccine & Infect Dis Div, Seattle, WA 98104 USA

来源：

CLINICAL CHEMISTRY | 2014年 / 60卷 / 05期

关键词：

DNA; BLOOD; HHV-6; QUANTITATION; REACTIVATION; INFECTION; THERAPY;

D O I：

10.1373/clinchem.2013.217240

中图分类号：

R446 [实验室诊断]; R-33 [实验医学、医学实验];

学科分类号：

1001 ;

摘要：

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. 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. 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. 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 stored plasma samples, facilitating retrospective analysis of the clinical significance of ciHHV-6. (C) 2014 American Association for Clinical Chemistry

引用

页码：765 / 772

页数：8

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