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Gene Expression Analysis by qPCR in Clinical Kidney Transplantation

  • Michael Eikmans
  • Jacqueline D. H. Anholts
  • Frans H. J. Claas
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1160)

Abstract

Patients with a kidney transplant may encounter chronic dysfunction of their graft. Once damage in the graft has established, therapeutic intervention is less efficient. Clinical parameters and morphologic evaluation of biopsies are used for determining diagnosis and prognosis of the patient. Quantitative polymerase chain reaction (qPCR) may be integrated in clinical practice to facilitate routine diagnostics, risk assessment with respect to graft outcome, and determination of the response to therapy by the patient. The success of qPCR assays is highly dependent on the adequacy of the methodological procedures performed. Here, we describe tips and tricks for processing patient material, RNA analysis, and qPCR primer design and gene expression analyses.

Key words

mRNA Transplant Kidney Diagnosis Prognosis 

Notes

Acknowledgement

Part of the results shown in Fig. 5 were derived from a project on B cells in kidney transplantation, initiated by Dr. Sebastiaan Heidt (Leiden University Medical Center, Department of Immunohematology).

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Michael Eikmans
    • 1
  • Jacqueline D. H. Anholts
    • 1
  • Frans H. J. Claas
    • 1
  1. 1.Department of Immunohematology and Blood TransfusionLeiden University Medical CenterLeidenThe Netherlands

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