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Reference: Karamitros, Timokratis, Paraskevis, D, Hatzakis, A et al., (2016). A contaminant-free assessment of Endogenous Retroviral RNA in human plasma. Scientific Reports, 6, 33598.Citable link to this page:

 

A contaminant-free assessment of Endogenous Retroviral RNA in human plasma

Abstract: Endogenous retroviruses (ERVs) comprise 6-8% of the human genome. HERVs are silenced in most normal tissues, up-regulated in stem cells and in placenta but also in cancer and HIV-1 infection. Crucially, there are conflicting reports on detecting HERV RNA in non-cellular clinical samples such as plasma that suggest the study of HERV RNA can be daunting. Indeed, we find that the use of real-time PCR in a quality assured clinical laboratory setting can be sensitive to low-level proviral contamination. We developed a mathematical model for low-level contamination that allowed us to design a laboratory protocol and standard operating procedures for robust measurement of HERV RNA. We focus on one family, HERV-K HML-2 (HK2) that has been most recently active even though they invaded our ancestral genomes almost 30 millions ago. We extensively validated our experimental design on a model cell culture system showing high sensitivity and specificity, totally eliminating the proviral contamination. We then tested 236 plasma samples from patients infected with HIV-1, HCV or HBV and found them to be negative. The study of HERV RNA for human translational studies should be performed with extensively validated protocols and standard operating procedures to control the widespread low-level human DNA contamination.

Publication status:PublishedPeer Review status:Peer reviewedVersion:Publisher's version Funder: Medical Research Council   Notes:This work is licensed under a Creative Commons Attribution 4.0 International License. The imagesor other third party material in this article are included in the article’s Creative Commons license,unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license,users will need to obtain permission from the license holder to reproduce the material. To view a copy of thislicense, visit http://creativecommons.org/licenses/by/4.0/

Bibliographic Details

Publisher: Nature Publishing Group

Publisher Website: http://www.nature.com/

Journal: Scientific Reportssee more from them

Publication Website: http://www.nature.com/srep/index.html

Volume: 6

Extent: 33598

Issue Date: 2016

pages:33598Identifiers

Doi: https://doi.org/10.1038/srep33598

Issn: 2045-2322

Uuid: uuid:5ff42044-57ec-433c-a020-96649a020545

Urn: uri:5ff42044-57ec-433c-a020-96649a020545

Pubs-id: pubs:645924 Item Description

Type: journal-article;

Language: eng

Version: Publisher's versionKeywords: Journal Article

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Autor: Karamitros, Timokratis - Oxford, MPLS, Zoology - - - Paraskevis, D - - - Hatzakis, A - - - Psichogiou, M - - - Elefsiniotis, I -

Fuente: https://ora.ox.ac.uk/objects/uuid:5ff42044-57ec-433c-a020-96649a020545



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