Journal of Molecular Medicine

, Volume 82, Issue 11, pp 750–755 | Cite as

Differential-display PCR of peripheral blood for biomarker discovery in chronic fatigue syndrome

  • Martin Steinau
  • Elizabeth R. Unger
  • Suzanne D. Vernon
  • James F. Jones
  • Mangalathu S. Rajeevan
Original Article

Abstract

We used differential-display PCR of peripheral blood mononuclear cells (PBMCs) to search for candidate biomarkers for chronic fatigue syndrome (CFS). PBMCs were collected from a subject with CFS and an age- and sex-matched control before and 24 h after exercise. RNA expression profiles were generated using 46 primer combinations, and the similarity between the individuals was striking. Differentially expressed bands were excised, reamplified, and sequenced, yielding 95 nonredundant sequences, of which 50 matched to known gene transcripts, 38 matched to genes with unknown functions, and 7 had no similarity to any database entry. Most (86%) of the differences between the two subjects were present at baseline. Differential expression of ten genes was verified by real-time reverse-transcription PCR: five (cystatin F, MHC class II, platelet factor 4, fetal brain expressed sequence tag, and perforin) were downregulated, and the remaining five genes (cathepsin B, DNA polymerase ε4, novel EST PBMC191MSt, heparanase precursor, and ORF2/L1 element) were upregulated in the subject with CFS. Many of these genes have known functions in defense and immunity, thus supporting prior suggestions of immune dysregulation in the pathogenesis of CFS. Differential-display PCR is a powerful tool for identification of candidate biomarkers. Investigation of these markers in samples from well-designed epidemiological studies of CFS will be required to determine the validity of these candidate biomarkers. The real-time reverse-transcription PCR assays that we developed for assay of these biomarkers will facilitate high-throughput testing of these additional samples.

Keywords

Chronic fatigue syndrome Gene expression profiling Differential display PCR Biomarkers Real-time reverse-transcription PCR 

Abbreviations

CFS

Chronic fatigue syndrome

Cp

Crossing point

DD-PCR

Differential-display polymerase chain reaction

EF

Elongation factor

EST

Expressed sequence tag

LINE

Long interspersed element

PBMC

Peripheral blood mononuclear cell

RT-PCR

Reverse-transcription polymerase chain reaction

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Martin Steinau
    • 1
  • Elizabeth R. Unger
    • 1
  • Suzanne D. Vernon
    • 1
  • James F. Jones
    • 1
  • Mangalathu S. Rajeevan
    • 1
  1. 1.Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and PreventionAtlantaUSA

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