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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 antibodies in CFS patients by Western blot (WB) analysis. Representative WB results for CFS cases from Wichita and Georgia identified after unblinding. Determination of MuLV specific reactivity is determined by comparison of observed seroreactivity to polytropic MuLV-infected HeLa antigens and uninfected HeLa antigens in upper and lower panels, respectively. Lanes 1 - 4 and 5 - 8 are plasma from CFS cases from the population based studies in Georgia and Wichita, respectively; lanes 9 - 12 are physician-referred CFS cases from the Georgia Registry study. MuLV positive and negative goat serum controls are labelled.
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Figure 3: Absence of XMRV antibodies in CFS patients by Western blot (WB) analysis. Representative WB results for CFS cases from Wichita and Georgia identified after unblinding. Determination of MuLV specific reactivity is determined by comparison of observed seroreactivity to polytropic MuLV-infected HeLa antigens and uninfected HeLa antigens in upper and lower panels, respectively. Lanes 1 - 4 and 5 - 8 are plasma from CFS cases from the population based studies in Georgia and Wichita, respectively; lanes 9 - 12 are physician-referred CFS cases from the Georgia Registry study. MuLV positive and negative goat serum controls are labelled.

Mentions: Plasma samples from 51 CFS cases and 53 healthy controls were diluted 1:50 and examined for seroreactivity to bands corresponding to Gag (p30 or p68/80) and/or Env (gp69/71 or p15E) proteins present in only the infected lysate and not the uninfected lysate. We also tested sera from 26 retrovirus-positive specimens (13 HTLV-1/2, seven HIV-1, and six dual HIV-1/HIV-2 seropositive patients) and observed no reactivity to XMRV proteins (data not shown) confirming a lack of cross-seroreactivity. In addition, we tested archived sera from 121 anonymous US blood donors; all were negative (data not shown). Plasma samples from the 51 CFS patients and 53 healthy controls all tested negative for XMRV antibodies in this assay. Plasma samples were not available from three healthy controls. Typical WB results of CFS persons are shown in Figure 3. Every plasma specimen demonstrated some level of background reactivity, but without evidence of specific reactivity to Gag and/or Env proteins (Figure 3). For example, plasma from a CFS person showed reactivity to two proteins about 65 and 69 kD in size in the infected cell lysate but reacted non-specifically to proteins of the same size in the uninfected antigen and was thus considered seronegative (lane 2 of Figure 3). There were no clear differences in nonspecific WB seroreactivity observed in healthy persons compared to persons with CFS (data not shown).


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 antibodies in CFS patients by Western blot (WB) analysis. Representative WB results for CFS cases from Wichita and Georgia identified after unblinding. Determination of MuLV specific reactivity is determined by comparison of observed seroreactivity to polytropic MuLV-infected HeLa antigens and uninfected HeLa antigens in upper and lower panels, respectively. Lanes 1 - 4 and 5 - 8 are plasma from CFS cases from the population based studies in Georgia and Wichita, respectively; lanes 9 - 12 are physician-referred CFS cases from the Georgia Registry study. MuLV positive and negative goat serum controls are labelled.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: Absence of XMRV antibodies in CFS patients by Western blot (WB) analysis. Representative WB results for CFS cases from Wichita and Georgia identified after unblinding. Determination of MuLV specific reactivity is determined by comparison of observed seroreactivity to polytropic MuLV-infected HeLa antigens and uninfected HeLa antigens in upper and lower panels, respectively. Lanes 1 - 4 and 5 - 8 are plasma from CFS cases from the population based studies in Georgia and Wichita, respectively; lanes 9 - 12 are physician-referred CFS cases from the Georgia Registry study. MuLV positive and negative goat serum controls are labelled.
Mentions: Plasma samples from 51 CFS cases and 53 healthy controls were diluted 1:50 and examined for seroreactivity to bands corresponding to Gag (p30 or p68/80) and/or Env (gp69/71 or p15E) proteins present in only the infected lysate and not the uninfected lysate. We also tested sera from 26 retrovirus-positive specimens (13 HTLV-1/2, seven HIV-1, and six dual HIV-1/HIV-2 seropositive patients) and observed no reactivity to XMRV proteins (data not shown) confirming a lack of cross-seroreactivity. In addition, we tested archived sera from 121 anonymous US blood donors; all were negative (data not shown). Plasma samples from the 51 CFS patients and 53 healthy controls all tested negative for XMRV antibodies in this assay. Plasma samples were not available from three healthy controls. Typical WB results of CFS persons are shown in Figure 3. Every plasma specimen demonstrated some level of background reactivity, but without evidence of specific reactivity to Gag and/or Env proteins (Figure 3). For example, plasma from a CFS person showed reactivity to two proteins about 65 and 69 kD in size in the infected cell lysate but reacted non-specifically to proteins of the same size in the uninfected antigen and was thus considered seronegative (lane 2 of Figure 3). There were no clear differences in nonspecific WB seroreactivity observed in healthy persons compared to persons with CFS (data not shown).

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