This was reposted from mecfsforums.com with the poster's permission:
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Abstract: O_11
XMRV in Chronic Fatigue Syndrome: A Pilot Study
M.R. Hanson, L.L. Lee, L. Lin, D.E. Bell, D. Ruppert, D. S. Bell
Cornell University, Dept of Molecular Biology and Genetics, Ithaca NY, USA; State University of New York, Dept of Medical Anthropology, Buffalo NY, USA; Cornell University, School of Operations Research and Information Engineering, Ithaca NY, USA; State University of New York, dept of Pediatrics, Buffalo NY, USA
Background:
In October 2009, Lombardi et all repoted detection of XMRV in blood of persons with CFS. Since then, four studies that performed PCR analysis on DNA from blood of CFS patients failed to detect XMRV. The present blinded study was undertaken to determine whether XMRV could be detected in peripheral blood mononuclear cells (PBMCs) from three small groups of subjects from a single geographic area.
Materials and methods:
The 30 adult subjects of this study were divided into three groups of ten persons each: severely ill with CFS, recovered from CFS, and a control group lacking a CFS diagnosis at any time. Inclusion did not require that they became ill in the time frame or in the exact location of a cluster of CFS in Lyndonville, NY. All patients in group 1 ("severe CFS") currently meet Fukuda criteria. The individuals in group 2 ("recovered CFS") had met Fukuda criteria for CFS at one time in the past, but presently considered themselves recovered. One patient in group 1 and seven in group 2 are considered part of the "Lyndonville outbreak". The ten persons included in group 3 "healthy, non-household contact controls" have never experienced a CFS-like illness, are healthy, and have never lived with persons with CFS. Average ages of the three groups are 42.2, 39, 49.8, respectively; and M:F ratio is 2:8 6:4, 2:8. All thirty subjects completed seven different clinical survey instruments including the SF-36. Blood was collected in EDTA tubes and PBMCs and plasma separated within 24 hours of draw. Plasma from some samples was incubated with LNCaP cells. The Invitrogen Superscript (R) VILO (TM) cDNA Synthesis Kit was used to produce cDNA from RNA isolated from PBMCs or LNCaP cells. Nested PCR with USB Hot-Start IT FideliTaq was performed using the Gag O-Gag l primers originally described by Urisman et al. PCR products were separated on gels and any bands of approximately 400 bp were sequenced and analyzed by BLAST for similarity to the XMRV gag gene. PCR with mouse COX2 primers was performed on all cDNA preparations to rule out mouse cell contamination.
Results:
XMRV gag sequences were detected in eight of the "severe CFS", three of the "recovered CFS" and in one of the controls. Plasma from six blood samples, five CFS and one control, was incubated with LNCaP cells; the six cultures were passaged six times. The five cultures found to exhibit XMRV gag sequences were those inoculated with CFS patient plastma. Although group 2 members described themselves as recovered, their scores on the SF-36 were significantly lower than the healthy control group, according to Hoteling's T2 test. Tukey's multiple comparison of means indicates that there are highly significant differences between the scores of the "severe" and controls on all 7 instruments.
Conclusions:
XMRV gag sequences were detected in a higher percentage of severe or recovered CFS cases (55%) compared to controls (10%) with one-sided p-value=0.0118. Human plasma samples from CFS cases contain infectious XMRV. Our results corroborate those of Lombardi et al.