http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0030889
Multiple Sources of Contamination in Samples from Patients Reported to
Have XMRV Infection
Mary F. Kearney1, Jonathan Spindler1, Ann Wiegand1, Wei Shao2,
Elizabeth M. Anderson1, Frank Maldarelli1, Francis W. Ruscetti3, John
W. Mellors4, Steve H. Hughes1, Stuart F. J. Le Grice1, John M. Coffin5
1 HIV Drug Resistance Program, National Cancer Institute, Frederick,
Maryland, United States of America, 2 Advanced Biomedical Computing
Center, SAIC, Frederick, Maryland, United States of America, 3
Laboratory of Experimental Immunology, Cancer and Inflammation
Program, National Cancer Institute, Frederick, Maryland, United States
of America, 4 Department of Medicine, University of Pittsburgh,
Pittsburgh, Pennsylvania, United States of America, 5 Department of
Molecular Biology and Microbiology, Tufts University, Boston,
Massachusetts, United States of America
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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