Skip to main content

Ultrasensitive PCR Detection of Tumor Cells in Myeloma

  • Protocol
PCR Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 226))

  • 7018 Accesses

Abstract

Chromosomal aberrations, such as translocations or inversions, described for a growing number of malignancies, are now widely used to detect tumor cells by polymerase chain reaction (PCR). However, in multiple myeloma (MM), no such ubiquitous PCR marker exists. Therefore, other means have been established to distinguish myeloma cells from normal cells. Because the plasma cells of a myeloma clone share an identical rearranged immunoglobulin gene sequence, it is possible to detect malignant cells with PCR primers specific for the VDJ rearrangement of the heavy chain of each myeloma clone. The sensitivity and specificity of this method, named allele-specific oligonucleotide (ASO) PCR, even with low proportions of malignant cells, has been proven (1).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 74.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Cremer, F. W., Kiel, K., Wallmeier, M., Goldschmidt, H., and Moos, M. (1997) A quantitative PCR assay for the detection of low amounts of malignant cells in multiple myeloma. Ann. Oncol. 8, 633–636.

    Article  PubMed  CAS  Google Scholar 

  2. Tonegawa, S. (1983) Somatic generation of antibody diversity. Nature 302, 575–581.

    Article  PubMed  CAS  Google Scholar 

  3. Walter, M. A., Surti, U., Hofker, M. H., and Cox, D. W. (1990) The physical organisation of the human immunoglobulin heavy chain gene complex. Eur. Mol. Biol. Org. J. 9, 3303–3313.

    CAS  Google Scholar 

  4. Bakkus, M. H., Heirman, C., Van Riet, I., Van Camp, B., and Thielemans, K. (1992) Evidence that multiple myeloma Ig heavy chain VDJ genes contain somatic mutations but show no intraclonal variation. Blood 80, 2326–2335.

    PubMed  CAS  Google Scholar 

  5. Aubin, J., Davi, F., Nguyen-Salomon, F., Leboeuf, D., Debert, C., Taher, M., et al. (1995) Description of a novel FR1 IgH PCR strategy and its comparison with three other strategies for the detection of clonality in B cell malignancies. Leukemia 9, 471–479.

    PubMed  CAS  Google Scholar 

  6. Brisco, M. J., Tan, L. W., Orsborn, A. M., and Morley, A. A. (1990) Development of a highly sensitive assay, based on the polymerase chain reaction, for rare B-lymphocyte clones in a polyclonal population. Br. J. Haematol. 75, 163–167.

    Article  PubMed  CAS  Google Scholar 

  7. Campbell, M. J., Zelenetz, A. D., Levy, S., Levy, R. (1992) Use of family specific leader region primers for PCR amplification of the human heavy chain variable region gene repertoire. Mol. Immunol. 29, 193–203.

    Article  PubMed  Google Scholar 

  8. Deane, M., McCarthy, K. P., Wiedemann, I. M., and Norton, J. D. (1991) An improved method for detection of B-lymphoid clonality by polymerase chain reaction. Br. J. Haematol. 5, 726–730.

    CAS  Google Scholar 

  9. Diss, T. C., Peng, H., Wotherspoon, A. C., Isaacson, P. G., and Pan, L. (1993) Detection of monoclonality in low-grade β-cell lymphoma using the polymerase chain reaction is dependent on primer selection and lymphoma type. J. Pathol. 169, 291–295.

    Article  PubMed  CAS  Google Scholar 

  10. Moore, G. E. and Kitamura, H. (1968) Cell line derived from patient with myeloma. N. Y. State J. Med. 68, 2054–2060.

    PubMed  CAS  Google Scholar 

  11. Klein, E., Klein, G., Nadkarmi, J. F., Nadkarmi, J. J., Vigzell, H., and Clifford, P. (1968) Surface IgM-k specificity on a Burkitt lymphoma cell in vivo and in derived culture lines. Cancer Res. 28, 1300–1310.

    PubMed  CAS  Google Scholar 

  12. Yamada, M., Wasserman, R., Reichard, B. A., Shane, S., Caton, A. J., and Rovera, G. (1991) Preferential utilization of specific immunoglobulin heavy chain diversity and joining segments in adult human peripheral blood B lymphocytes. J. Exp. Med. 173, 395–407.

    Article  PubMed  CAS  Google Scholar 

  13. Brisco, M. J., Condon, J., Sykes, P. J., Neoh, S. H., and Morley, A. A. (1991) Detection and quantitation of neoplastic cells in acute lymphoblastic leukaemia, by use of the polymerase chain reaction. Br. J. Haematol. 79, 211–217.

    Article  PubMed  CAS  Google Scholar 

  14. Wiesner, R. J. (1992) Direct quantitation of picomolar concentrations of mRNAs by mathematical analysis of a reverse transcription/exponential polymerase chain reaction assay. Nucleic Acids Res. 51, 5863–5864.

    Article  Google Scholar 

  15. Taswell, C. (1981) Limiting dilution assays for the determination of immunocompetent cell frequencies. J. Immunol. 126, 1614–1619.

    PubMed  CAS  Google Scholar 

  16. Cremer, F. W., Dada, R., Kiel, K., Ehrbrecht, E., Goldschmidt, H., and Moos, M. (1998) Quantitative PCR monitoring of the effect of double high-dose therapy on the tumor load of patients with multiple myeloma. Ann. Hematol. 77(Suppl. II), 182 (abstract 722).

    Google Scholar 

  17. Vescio, R. A., Han, E. J., Schiller, G. J., Lee, J. C., Wu, C. H., Cao, J., et al. (1996) Quantitative comparison of multiple myeloma tumor contamination in bone marrow harvest and leukapheresis autografts. Bone Marrow Transplant. 18, 103–110.

    PubMed  CAS  Google Scholar 

  18. Cremer, F. W., Kiel, K., Wallmeier, M., Haas, R., Goldschmidt, H., and Moos, M. (1998) Leukapheresis products in multiple myeloma: lower tumor load after mobilization with cyclophosphamide plus granulocyte colony-stimulating factor (G-CSF) compared with G-CSF alone. Exp. Hematol. 26, 969–975.

    PubMed  CAS  Google Scholar 

  19. Kiel, K., Cremer, F. W., Ehrbrecht, E., Wallmeier, M., Hegenbart, U., Goldschmidt, H., et al. (1998) First and second apheresis in patients with multiple myeloma: no differences in tumor load and hematopoietic stem cell yield. Bone Marrow Transplant. 21, 1109–1115.

    Article  PubMed  CAS  Google Scholar 

  20. Kiel, K., Cremer, F. W., Rottenburger, C., Kallmeyer, C., Ehrbrecht, E., Atzberger, A., et al. (1998) Circulating tumor cells in patients with multiple myeloma in the course of high-dose therapy. Ann. Hematol. 77(Suppl. II), 182 (abstract 724).

    Google Scholar 

  21. Rottenburger, C., Kiel, K., Cremer, F. W., B″sing, T., Atzberger, A., Moldenhauer, G., et al. (1998) No differences in the tumor load in the CD19+ and CD20+ cell fractions of peripheral blood from patients with multiple myeloma post high-dose therapy and with peripheral blood stem cell support. Ann. Hematol. 77(Suppl. II), 184 (abstract 731).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2003 Humana Press Inc.

About this protocol

Cite this protocol

Cremer, F.W., Moos, M. (2003). Ultrasensitive PCR Detection of Tumor Cells in Myeloma. In: Bartlett, J.M.S., Stirling, D. (eds) PCR Protocols. Methods in Molecular Biology™, vol 226. Humana Press. https://doi.org/10.1385/1-59259-384-4:185

Download citation

  • DOI: https://doi.org/10.1385/1-59259-384-4:185

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-642-0

  • Online ISBN: 978-1-59259-384-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics