2016
DOI: 10.1016/j.jcv.2016.02.016
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Efficient identification of inherited chromosomally integrated human herpesvirus 6 using specimen pooling

Abstract: Background Human herpesvirus 6 (HHV-6) has a unique ability to integrate into chromosomal telomeres. Vertical transmission via germ cell integration results in offspring with inherited chromosomally integrated (ci)HHV-6 in all nucleated cells, affecting ~1% of the population. Objectives Inherited ciHHV-6 may be a direct or indirect mediator of human disease, but efficient identification of affected individuals is a fundamental roadblock to larger studies exploring the clinical importance of this condition. … Show more

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Cited by 22 publications
(20 citation statements)
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“…13 We used droplet digital (dd)PCR to determine the ratio of HHV-6 DNA to human cells (using human ribonuclease P DNA [RPP30], a reference gene for cell count). 13,16,19 A ratio of 1 6 0.07 was considered positive for iciHHV-6 and distinguished between iciHHV-6 and naturally acquired infection. This assay was validated against a fluorescence in situ hybridizationconfirmed iciHHV-6 cell line.…”
Section: Laboratory Assaysmentioning
confidence: 99%
See 1 more Smart Citation
“…13 We used droplet digital (dd)PCR to determine the ratio of HHV-6 DNA to human cells (using human ribonuclease P DNA [RPP30], a reference gene for cell count). 13,16,19 A ratio of 1 6 0.07 was considered positive for iciHHV-6 and distinguished between iciHHV-6 and naturally acquired infection. This assay was validated against a fluorescence in situ hybridizationconfirmed iciHHV-6 cell line.…”
Section: Laboratory Assaysmentioning
confidence: 99%
“…16 This approach leveraged the relatively low population prevalence of iciHHV-6 and high individual viral burden among affected individuals to efficiently identify iciHHV-6. Briefly, approximately 5 mg DNA per sample was aliquoted in 5 mL with the addition of 20 mL buffer AE (Qiagen, CA) (0.2 mg/mL).…”
Section: Screening Algorithm For Icihhv-6mentioning
confidence: 99%
“…6 Since this seminal work, pooling techniques have been successfully implemented to screen for many other infectious diseases, including HIV, hepatitis B virus (HBV), hepatitis C virus (HCV), 7 influenza, 8 and herpes. 9 Besides disease screening, many other areas, including genetics, 10 veterinary science, 11 medical entomology, 12 blood safety, 13 and drug discovery, 14 have also used the method of pooling. Statistical research in group testing primarily focused on improving the diagnostic accuracy and cost-saving ability of a pooling protocol 15 or estimating individual-level characteristics from pooled testing data.…”
Section: Literature Reviewmentioning
confidence: 99%
“…Pooled testing (also known as group testing) was initially proposed to screen for syphilis among War World II American army recruits . Since this seminal work, pooling techniques have been successfully implemented to screen for many other infectious diseases, including HIV, hepatitis B virus (HBV), hepatitis C virus (HCV), influenza, and herpes . Besides disease screening, many other areas, including genetics, veterinary science, medical entomology, blood safety, and drug discovery, have also used the method of pooling.…”
Section: Introductionmentioning
confidence: 99%
“…In addition, the rise in HHV‐6 copies paralleled engraftment and rising WBC. A diagnosis of ciHHV‐6 was confirmed by a whole blood quantitative droplet digital PCR assay (ddPCR; Molecular Virology, University of Washington, Seattle, WA, USA) on Day +27 of the second syngeneic HCT using a previously described assay . Briefly, ddPCR provides a precise calculation of the number of HHV‐6 DNA copies per cell; this ratio is expected to be 1 in patients with ciHHV‐6.…”
Section: Case Reportmentioning
confidence: 99%