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Multiple sources of contamination in samples from patients reported to have XMRV infection

Kearney, MF and Spindler, J and Wiegand, A and Shao, W and Anderson, EM and Maldarelli, F and Ruscetti, FW and Mellors, JW and Hughes, SH and Le Grice, SFJ and Coffin, JM (2012) Multiple sources of contamination in samples from patients reported to have XMRV infection. PLoS ONE, 7 (2).

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Abstract

Xenotropic murine leukemia virus (MLV)-related retrovirus (XMRV) was reported to be associated with prostate cancer by Urisman, et al. in 2006 and chronic fatigue syndrome (CFS) by Lombardi, et al. in 2009. To investigate this association, we independently evaluated plasma samples from 4 patients with CFS reported by Lombardi, et al. to have XMRV infection and from 5 healthy controls reported to be XMRV uninfected. We also analyzed viral sequences obtained from supernatants of cell cultures found to contain XMRV after coculture with 9 clinical samples from 8 patients. A qPCR assay capable of distinguishing XMRV from endogenous MLVs showed that the viral sequences detected in the CFS patient plasma behaved like endogenous MLVs and not XMRV. Single-genome sequences (N = 89) from CFS patient plasma were indistinguishable from endogenous MLVs found in the mouse genome that are distinct from XMRV. By contrast, XMRV sequences were detected by qPCR in 2 of the 5 plasma samples from healthy controls (sequencing of the qPCR product confirmed XMRV not MLV). Single-genome sequences (N = 234) from the 9 culture supernatants reportedly positive for XMRV were indistinguishable from XMRV sequences obtained from 22Rv1 and XMRV-contaminated 293T cell-lines. These results indicate that MLV DNA detected in the plasma samples from CFS patients evaluated in this study was from contaminating mouse genomic DNA and that XMRV detected in plasma samples from healthy controls and in cultures of patient samples was due to cross-contamination with XMRV (virus or nucleic acid).

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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Kearney, MF
Spindler, J
Wiegand, A
Shao, W
Anderson, EM
Maldarelli, F
Ruscetti, FW
Mellors, JWjwm1@pitt.eduJWM1
Hughes, SH
Le Grice, SFJ
Coffin, JM
Contributors:
ContributionContributors NameEmailPitt UsernameORCID
EditorApetrei, Cristianapetreic@pitt.eduAPETREICUNSPECIFIED
Date: 20 February 2012
Date Type: Publication
Journal or Publication Title: PLoS ONE
Volume: 7
Number: 2
DOI or Unique Handle: 10.1371/journal.pone.0030889
Refereed: Yes
MeSH Headings: Animals; Base Sequence; Cell Line, Tumor; Coculture Techniques; DNA Contamination; DNA, Viral--blood; DNA, Viral--genetics; Fatigue Syndrome, Chronic--blood; Fatigue Syndrome, Chronic--genetics; Fatigue Syndrome, Chronic--virology; Female; Gene Products, env--genetics; Genetic Variation; Humans; Male; Mice; Molecular Sequence Data; Phylogeny; Polymerase Chain Reaction; Retroviridae Infections--blood; Retroviridae Infections--genetics; Retroviridae Infections--virology; Subcellular Fractions--metabolism; Xenotropic murine leukemia virus-related virus--genetics; Xenotropic murine leukemia virus-related virus--isolation & purification
Other ID: NLM PMC3282701
PubMed Central ID: PMC3282701
PubMed ID: 22363509
Date Deposited: 13 Sep 2012 18:02
Last Modified: 02 Feb 2019 16:57
URI: http://d-scholarship.pitt.edu/id/eprint/14141

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