Premium
Selection of reference genes for quantitative polymerase chain reaction studies in purified B cells from B cell chronic lymphocytic leukaemia patients
Author(s) -
Valceckiene Vilma,
Kontenyte Rima,
Jakubauskas Arturas,
Griskevicius Laimonas
Publication year - 2010
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.2010.08363.x
Subject(s) - reference genes , biology , gene , polymerase chain reaction , cd19 , gene expression , real time polymerase chain reaction , glyceraldehyde 3 phosphate dehydrogenase , b cell , microbiology and biotechnology , gene expression profiling , reverse transcription polymerase chain reaction , chronic lymphocytic leukemia , immunology , genetics , cell , leukemia , antibody
Summary The clinical heterogeneity of B‐cell chronic lymphocytic leukaemia (B‐CLL) makes it necessary to identify potent prognostic indicators to predict individual clinical course and select risk‐adapted therapy. In recent years, numerous gene expression models have been suggested as prognostic factors of B‐CLL. Today, quantitative polymerase chain reaction (qPCR) is a preferred method for rapid quantification of gene expression and validation of microarray data. The reliability of qPCR data is highly dependent on the use of appropriate reference genes for normalization. To date, no validated reference genes have been reported for the normalization of gene expression in B‐CLL. Therefore, the present study was conducted to identify suitable reference genes for gene expression studies in CD19 + B cells isolated from B‐CLL patients’ peripheral blood . The stability of ACTB , B2M , GAPDH , GUSB , HMBS , HPRT1 , MRPL19 , TBP and UBC genes was determined by three different descriptive statistics, geNorm, NormFinder and BestKeeper‐1, which produced highly comparable results. Based on our results, B2M , HPRT1 , and GUSB were found to be the most suitable reference genes for qPCR studies in B‐CLL patients’ peripheral blood B cells.