Abstract
BackgroundXMRV 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 FindingsTesting 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/SignificanceMethodologies 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.
Highlights
A novel retrovirus, xenotropic murine leukaemia virus-related virus (XMRV), probably of mouse origin, was identified in nucleic acid (NA) extracted from human prostate cancers using a multi-virus array [1]
In initial experiments designed to test methodology for screening the blood supply, we examined 80 cDNAs from normal blood donors using the modified XMRV gag TaqMan protocol
When the Invitrogen Platinum Taq (IPT) reagents were substituted with Applied Biosystems Taq Gold LD (ABTG) reagents in a second set of experiments, re-amplification of 8 cDNAs, including 7C and 9C, using the nested gag primers failed to generate any detectable products
Summary
A novel retrovirus, xenotropic murine leukaemia virus-related virus (XMRV), probably of mouse origin, was identified in nucleic acid (NA) extracted from human prostate cancers using a multi-virus array [1]. Late in 2009 Lombardi and colleagues reported the presence of XMRV DNA in peripheral blood leucocytes from 67% of patients with chronic fatigue syndrome (CFS) and 3.7% of healthy controls [3]. 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. We decided to investigate blood donors for evidence of XMRV/pMLV
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