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

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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.

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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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Acknowledgements

The expert technical assistance of Ms. Dana Taysavang and Dr. Dominique Rollin is gratefully acknowledged. The authors thank Dr. William C. Reeves for critical appraisal of this work and Ms. Bristol Sorensen and Joanne Streib for subject enrollment and specimen processing. J.F.J. was affiliated with the National Jewish Medical and Research Center, Denver, Colo., USA, during part of the study. This study adhered to human experimentation guidelines of the United States Department of Health and Human Services and was reviewed and approved by the Institutional Review Boards of the Centers for Disease Control and Prevention and National Jewish Medical Center. All participants were volunteers who gave informed consent.

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Authors and Affiliations

  1. Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Rd., MSG-41, Atlanta, GA, 30333, USA

    Martin Steinau, Elizabeth R. Unger, Suzanne D. Vernon, James F. Jones & Mangalathu S. Rajeevan

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  1. Martin Steinau
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  2. Elizabeth R. Unger
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  3. Suzanne D. Vernon
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  4. James F. Jones
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  5. Mangalathu S. Rajeevan
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Correspondence to Mangalathu S. Rajeevan.

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Steinau, M., Unger, E.R., Vernon, S.D. et al. Differential-display PCR of peripheral blood for biomarker discovery in chronic fatigue syndrome. J Mol Med 82, 750–755 (2004). https://doi.org/10.1007/s00109-004-0586-4

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  • DOI: https://doi.org/10.1007/s00109-004-0586-4

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