title: Gene expression analysis by qPCR in clinical kidney transplantation.
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.
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Albinusdreef 2, Leiden, 2333 ZA, The Netherlands, m.eikmans@lumc.nl.
authors.forename: Michael
authors.initials: M
authors.lastname: Eikmans
authors.forename: Jacqueline D H
authors.initials: JD
authors.lastname: Anholts
authors.forename: Frans H J
authors.initials: FH
authors.lastname: Claas
mesh_terms: D001773:Blood Cells / Q000378:metabolism; D005615:Freezing; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D016030:Kidney Transplantation; D016133:Polymerase Chain Reaction / Q000379:methods*