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Absence of evidence of xenotropic murine leukemia virus-related virus infection in persons with chronic fatigue syndrome and healthy controls in the United States.

Switzer WM, Jia H, Hohn O, Zheng H, Tang S, Shankar A, Bannert N, Simmons G, Hendry RM, Falkenberg VR, Reeves WC, Heneine W - Retrovirology (2010)

Bottom Line: Additional blinded screening of the 51 cases and 53 controls at the Robert Koch Institute using an ELISA employing recombinant Gag and Env XMRV proteins identified weak seroreactivity in one CFS case and a healthy control, which was not confirmed by immunofluorescence.DNA specimens from 50 CFS patients and 56 controls and 41 US blood donors were all PCR-negative.These data do not support an association of XMRV with CFS.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory Branch, Division of HIV/AIDS Prevention, National Center for HIV/AIDS, Viral Hepatitis, STD, and TB Prevention, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA. bswitzer@cdc.gov

ABSTRACT

Background: XMRV, a xenotropic murine leukemia virus (MuLV)-related virus, was recently identified by PCR testing in 67% of persons with chronic fatigue syndrome (CFS) and in 3.7% of healthy persons from the United States. To investigate the association of XMRV with CFS we tested blood specimens from 51 persons with CFS and 56 healthy persons from the US for evidence of XMRV infection by using serologic and molecular assays. Blinded PCR and serologic testing were performed at the US Centers for Disease Control and Prevention (CDC) and at two additional laboratories.

Results: Archived blood specimens were tested from persons with CFS defined by the 1994 international research case definition and matched healthy controls from Wichita, Kansas and metropolitan, urban, and rural Georgia populations. Serologic testing at CDC utilized a Western blot (WB) assay that showed excellent sensitivity to MuLV and XMRV polyclonal or monoclonal antibodies, and no reactivity on sera from 121 US blood donors or 26 HTLV-and HIV-infected sera. Plasma from 51 CFS cases and plasma from 53 controls were all WB negative. Additional blinded screening of the 51 cases and 53 controls at the Robert Koch Institute using an ELISA employing recombinant Gag and Env XMRV proteins identified weak seroreactivity in one CFS case and a healthy control, which was not confirmed by immunofluorescence. PCR testing at CDC employed a gag and a pol nested PCR assay with a detection threshold of 10 copies in 1 ug of human DNA. DNA specimens from 50 CFS patients and 56 controls and 41 US blood donors were all PCR-negative. Blinded testing by a second nested gag PCR assay at the Blood Systems Research Institute was also negative for DNA specimens from the 50 CFS cases and 56 controls.

Conclusions: We did not find any evidence of infection with XMRV in our U.S. study population of CFS patients or healthy controls by using multiple molecular and serologic assays. These data do not support an association of XMRV with CFS.

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Absence of XMRV polymerase (pol) sequences in CFS patients. A. Representative nested pol PCR results using PBMC DNA specimens from persons with CFS identified after unblinding. Lanes 1 - 5, 6 - 10, and 11 - 14 are results for persons with CFS from Wichita, Georgia, and the Georgia registry studies, respectively; lanes 15 and 16, water only controls; lane 17, negative human PBMC DNA control; lanes 18 and 19, assay sensitivity controls consisting of 101 and 103 copies of XMRV VP62 plasmid DNA diluted in a background of 1 ug of human PBMC DNA, respectively. B. Semi-quantitative β-actin PCR results for PBMC DNA specimens above in lanes 1 - 14; lane 15, water control; lanes 16 - 19, 10-fold dilutions of blood donor PBMC DNA starting at 0.1 ug as a positive assay control.
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Figure 5: Absence of XMRV polymerase (pol) sequences in CFS patients. A. Representative nested pol PCR results using PBMC DNA specimens from persons with CFS identified after unblinding. Lanes 1 - 5, 6 - 10, and 11 - 14 are results for persons with CFS from Wichita, Georgia, and the Georgia registry studies, respectively; lanes 15 and 16, water only controls; lane 17, negative human PBMC DNA control; lanes 18 and 19, assay sensitivity controls consisting of 101 and 103 copies of XMRV VP62 plasmid DNA diluted in a background of 1 ug of human PBMC DNA, respectively. B. Semi-quantitative β-actin PCR results for PBMC DNA specimens above in lanes 1 - 14; lane 15, water control; lanes 16 - 19, 10-fold dilutions of blood donor PBMC DNA starting at 0.1 ug as a positive assay control.

Mentions: PCR testing of β-actin sequences was positive for all clinical specimens confirming the integrity of the DNA and an absence of PCR inhibitors. Representative β-actin PCR results are shown in Figure 5. Subsequent XMRV testing showed that XMRV gag and pol sequences were not detected in 1 ug of PBMC (n = 31) or whole blood (n = 19) DNA from the CFS patients or in 1 ug PBMC DNA from the 56 healthy controls. A representative Southern blot of the XMRV pol PCR testing of persons with CFS is shown in Figure 5. Matching DNA was not available from one CFS case.


Absence of evidence of xenotropic murine leukemia virus-related virus infection in persons with chronic fatigue syndrome and healthy controls in the United States.

Switzer WM, Jia H, Hohn O, Zheng H, Tang S, Shankar A, Bannert N, Simmons G, Hendry RM, Falkenberg VR, Reeves WC, Heneine W - Retrovirology (2010)

Absence of XMRV polymerase (pol) sequences in CFS patients. A. Representative nested pol PCR results using PBMC DNA specimens from persons with CFS identified after unblinding. Lanes 1 - 5, 6 - 10, and 11 - 14 are results for persons with CFS from Wichita, Georgia, and the Georgia registry studies, respectively; lanes 15 and 16, water only controls; lane 17, negative human PBMC DNA control; lanes 18 and 19, assay sensitivity controls consisting of 101 and 103 copies of XMRV VP62 plasmid DNA diluted in a background of 1 ug of human PBMC DNA, respectively. B. Semi-quantitative β-actin PCR results for PBMC DNA specimens above in lanes 1 - 14; lane 15, water control; lanes 16 - 19, 10-fold dilutions of blood donor PBMC DNA starting at 0.1 ug as a positive assay control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2908559&req=5

Figure 5: Absence of XMRV polymerase (pol) sequences in CFS patients. A. Representative nested pol PCR results using PBMC DNA specimens from persons with CFS identified after unblinding. Lanes 1 - 5, 6 - 10, and 11 - 14 are results for persons with CFS from Wichita, Georgia, and the Georgia registry studies, respectively; lanes 15 and 16, water only controls; lane 17, negative human PBMC DNA control; lanes 18 and 19, assay sensitivity controls consisting of 101 and 103 copies of XMRV VP62 plasmid DNA diluted in a background of 1 ug of human PBMC DNA, respectively. B. Semi-quantitative β-actin PCR results for PBMC DNA specimens above in lanes 1 - 14; lane 15, water control; lanes 16 - 19, 10-fold dilutions of blood donor PBMC DNA starting at 0.1 ug as a positive assay control.
Mentions: PCR testing of β-actin sequences was positive for all clinical specimens confirming the integrity of the DNA and an absence of PCR inhibitors. Representative β-actin PCR results are shown in Figure 5. Subsequent XMRV testing showed that XMRV gag and pol sequences were not detected in 1 ug of PBMC (n = 31) or whole blood (n = 19) DNA from the CFS patients or in 1 ug PBMC DNA from the 56 healthy controls. A representative Southern blot of the XMRV pol PCR testing of persons with CFS is shown in Figure 5. Matching DNA was not available from one CFS case.

Bottom Line: Additional blinded screening of the 51 cases and 53 controls at the Robert Koch Institute using an ELISA employing recombinant Gag and Env XMRV proteins identified weak seroreactivity in one CFS case and a healthy control, which was not confirmed by immunofluorescence.DNA specimens from 50 CFS patients and 56 controls and 41 US blood donors were all PCR-negative.These data do not support an association of XMRV with CFS.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory Branch, Division of HIV/AIDS Prevention, National Center for HIV/AIDS, Viral Hepatitis, STD, and TB Prevention, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA. bswitzer@cdc.gov

ABSTRACT

Background: XMRV, a xenotropic murine leukemia virus (MuLV)-related virus, was recently identified by PCR testing in 67% of persons with chronic fatigue syndrome (CFS) and in 3.7% of healthy persons from the United States. To investigate the association of XMRV with CFS we tested blood specimens from 51 persons with CFS and 56 healthy persons from the US for evidence of XMRV infection by using serologic and molecular assays. Blinded PCR and serologic testing were performed at the US Centers for Disease Control and Prevention (CDC) and at two additional laboratories.

Results: Archived blood specimens were tested from persons with CFS defined by the 1994 international research case definition and matched healthy controls from Wichita, Kansas and metropolitan, urban, and rural Georgia populations. Serologic testing at CDC utilized a Western blot (WB) assay that showed excellent sensitivity to MuLV and XMRV polyclonal or monoclonal antibodies, and no reactivity on sera from 121 US blood donors or 26 HTLV-and HIV-infected sera. Plasma from 51 CFS cases and plasma from 53 controls were all WB negative. Additional blinded screening of the 51 cases and 53 controls at the Robert Koch Institute using an ELISA employing recombinant Gag and Env XMRV proteins identified weak seroreactivity in one CFS case and a healthy control, which was not confirmed by immunofluorescence. PCR testing at CDC employed a gag and a pol nested PCR assay with a detection threshold of 10 copies in 1 ug of human DNA. DNA specimens from 50 CFS patients and 56 controls and 41 US blood donors were all PCR-negative. Blinded testing by a second nested gag PCR assay at the Blood Systems Research Institute was also negative for DNA specimens from the 50 CFS cases and 56 controls.

Conclusions: We did not find any evidence of infection with XMRV in our U.S. study population of CFS patients or healthy controls by using multiple molecular and serologic assays. These data do not support an association of XMRV with CFS.

Show MeSH
Related in: MedlinePlus