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. 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 provides excellent sensitivity and specificity using stored plasma samples, facilitating retrospective analysis of the clinical significance of ciHHV-6.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Sedlak RH,Cook L,Huang ML,Magaret A,Zerr DM,Boeckh M,Jerome KRdoi
10.1373/clinchem.2013.217240subject
Has Abstractpub_date
2014-05-01 00:00:00pages
765-72issue
5eissn
0009-9147issn
1530-8561pii
clinchem.2013.217240journal_volume
60pub_type
杂志文章abstract：BACKGROUND:The multiple endocrine neoplasia 2 (MEN 2) syndromes [MEN 2A, MEN 2B, and familial medullary thyroid carcinoma (FMTC)] are caused by germline mutations of the RET protooncogene. Because 85% of MEN 2A patients and 30% of FMTC patients have mutations at codon 634, the recommended molecular analyses begin at ex...
journal_title：Clinical chemistry
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更新日期：2001-11-01 00:00:00
abstract：:We describe a completely automated enzymic system for measuring total cholesterol in serum. All reagents are contained in an analytical test pack and the test is performed on Du Pont's Automatic Clinical Analyzer (aca), which mixes the sample (20 microliter) and reagents and performs the necessary absorbance measureme...
journal_title：Clinical chemistry
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journal_title：Clinical chemistry
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journal_title：Clinical chemistry
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journal_title：Clinical chemistry
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更新日期：1998-07-01 00:00:00
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journal_title：Clinical chemistry
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更新日期：1999-12-01 00:00:00
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journal_title：Clinical chemistry
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更新日期：2002-03-01 00:00:00
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journal_title：Clinical chemistry
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更新日期：1984-08-01 00:00:00
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journal_title：Clinical chemistry
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doi：
更新日期：1986-03-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1988-04-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1994-11-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1986-10-01 00:00:00
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doi：
更新日期：1989-04-01 00:00:00
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doi：10.1373/clinchem.2004.037606
更新日期：2005-02-01 00:00:00
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journal_title：Clinical chemistry
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doi：
更新日期：1987-04-01 00:00:00
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journal_title：Clinical chemistry
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doi：
更新日期：1991-11-01 00:00:00
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更新日期：1992-06-01 00:00:00
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更新日期：1996-07-01 00:00:00
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doi：
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doi：
更新日期：1988-09-01 00:00:00
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更新日期：1984-12-01 00:00:00
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journal_title：Clinical chemistry
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doi：
更新日期：1979-02-01 00:00:00
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doi：
更新日期：1987-01-01 00:00:00
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journal_title：Clinical chemistry
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doi：
更新日期：1981-04-01 00:00:00
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doi：
更新日期：2001-02-01 00:00:00
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