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PCR Master Mixes Harbour Murine DNA Sequences. Caveat Emptor!

Tuke, PW; Tettmar, KI; Tamuri, A; Stoye, JP; Tedder, RS; (2011) PCR Master Mixes Harbour Murine DNA Sequences. Caveat Emptor! PLoS One , 6 (5) , Article e19953. 10.1371/journal.pone.0019953. Green open access

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Abstract

Background XMRV is the most recently described retrovirus to be found in Man, firstly in patients with prostate cancer (PC) and secondly in 67% of patients with chronic fatigue syndrome (CFS) and 3.7% of controls. Both disease associations remain contentious. Indeed, a recent publication has concluded that “XMRV is unlikely to be a human pathogen”. Subsequently related but different polytropic MLV (pMLV) sequences were also reported from the blood of 86.5% of patients with CFS. and 6.8% of controls. Consequently we decided to investigate blood donors for evidence of XMRV/pMLV. Methodology/Principal Findings Testing of cDNA prepared from the whole blood of 80 random blood donors, generated gag PCR signals from two samples (7C and 9C). These had previously tested negative for XMRV by two other PCR based techniques. To test whether the PCR mix was the source of these sequences 88 replicates of water were amplified using Invitrogen Platinum Taq (IPT) and Applied Biosystems Taq Gold LD (ABTG). Four gag sequences (2D, 3F, 7H, 12C) were generated with the IPT, a further sequence (12D) by ABTG re-amplification of an IPT first round product. Sequence comparisons revealed remarkable similarities between these sequences, endogeous MLVs and the pMLV sequences reported in patients with CFS. Conclusions/Significance Methodologies for the detection of viruses highly homologous to endogenous murine viruses require special caution as the very reagents used in the detection process can be a source of contamination and at a level where it is not immediately apparent. It is suggested that such contamination is likely to explain the apparent presence of pMLV in CFS.

Type: Article
Title: PCR Master Mixes Harbour Murine DNA Sequences. Caveat Emptor!
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1371/journal.pone.0019953
Publisher version: http://dx.doi.org/10.1371/journal.pone.0019953
Language: English
Additional information: © 2011 Tuke et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. PMCID: PMC3102076
Keywords: Animals, DNA Contamination, Humans, Indicators and Reagents, Mice, Polymerase Chain Reaction, RNA, Viral, Xenotropic murine leukemia virus-related virus
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
URI: https://discovery.ucl.ac.uk/id/eprint/1415492
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