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Summary

Contamination in PCR reagents may explain the presence of xenotropic murine leukemia virus (XMRV) and polytropic MLV (pMLV) in chronic fatigue syndrome (CFS) patients. This study investigated blood donors for XMRV/pMLV, finding reagent contamination was a likely source.

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Area of Science:

  • Virology
  • Molecular Biology
  • Immunology

Background:

  • Xenotropic murine leukemia virus (XMRV) detection in humans, particularly in prostate cancer and chronic fatigue syndrome (CFS) patients, remains controversial.
  • Previous studies reported XMRV and related polytropic MLV (pMLV) sequences in CFS patients, but these findings are debated.
  • The possibility of XMRV/pMLV being a human pathogen is questioned due to recent conflicting evidence.

Purpose of the Study:

  • To investigate the presence of XMRV and pMLV in blood donors.
  • To determine if PCR reagents could be a source of XMRV/pMLV sequences.
  • To assess the validity of previous XMRV/pMLV findings in CFS patients.

Main Methods:

  • Testing of cDNA from 80 random blood donors for XMRV/pMLV gag sequences using PCR.
  • Amplification of water replicates with two different Taq polymerases (Invitrogen Platinum Taq and Applied Biosystems Taq Gold LD) to check for reagent contamination.
  • Sequence comparison of amplified products with known MLV sequences.

Main Results:

  • Two blood donor samples showed gag PCR signals, despite negative results from other PCR techniques.
  • PCR reagent contamination was confirmed, with gag sequences generated from water replicates using both tested Taq polymerases.
  • Amplified sequences exhibited high similarity to endogenous murine leukemia viruses (MLVs) and previously reported pMLV sequences from CFS patients.

Conclusions:

  • Detection methodologies for viruses homologous to endogenous murine viruses require stringent precautions against reagent contamination.
  • Contamination of PCR reagents is a likely explanation for the reported presence of pMLV in CFS patients.
  • The findings highlight the critical importance of rigorous controls in molecular detection assays to avoid false-positive results.