WT1 RQ Kit

Reagent kit for quantitative determination of the WT1 gene expression levels in clinical material using polymerase chain reaction (PCR) using real-time hybridization-fluorescence detection.

 This technique can be used to assess probability of relapses in patients with acute leukemias.

The kit of reagents is designed to perform PCR assay in a quantitative format for 24 clinical samples in duplicate (a total of 132 PCR reactions, including controls). The test system is compatible to different PCR cyclers: "iQ5" (Bio-Rad); "CFX96" (Bio-Rad); "ABIPrism" (ThermoFisher Scientific); "Lightcycler" (Roche); "Rotor-Gene" 3000/6000 (Corbett Research/Qiagen).

Additional information about the molecular marker:

To date, molecular biological monitoring of minimal residual disease (MRD) in patients with AML is the most important technique when assessing effectiveness of anticancer therapy. However, feasibility of this approach is limited by the presence of chimeric marker genes in leukemia cells of individual patients detectable by quantitative real-time PCR, e.g., PML-RARA, CBFB-MYH11, AML1-ETO (RUNX1-RUNX1T1), or specific mutations in FLT3 and NPM1 genes. About a half of AML patients do not express such molecular genetic markers. Just for this group of patients, a search for alternative approaches to MRD monitoring is a key factor in assessing the effectiveness of therapy. One such approach is based on real-time quantitative PCR to detect gene transcripts which are over-expressed in leukemia cells compared with normal bone marrow and peripheral blood cells. WT1 (Wilms tumor 1) gene expression is the most striking example of such leukemia markers.

The WT1 gene is located at 11p13 and encodes a transcription factor of the "zinc fingers" type, which performs a broad range of functions in the cell, depending on the splicing variant of mRNA produced. Among the main WT1 functions during normal hematopoiesis, is the induction of stem cell differentiation to the myelo-monocytic lineage (1). WT1 is considered a normal tumor suppressor being responsible for mRNA maturation.

Overexpression of the WT1 gene has been found in a number of malignant blood diseases, including AML (2). High expression of WT1 occurs in more than half of AML patients, being a poor prognostic factor of the disease and high relapse risk (3). Since real-time quantitative PCR can detect distinct differences in WT1 transcript levels between normal and leukemia cells, the WT1 expression may be a useful marker of MRD in patients who do not have major chromosomal abnormalities or other mutational features of leukemic lineage cells.

Related publications:

     1Yang L, Han Y, Suarez Saiz F, Minden MD. A tumor suppressor and oncogene: the WT1 story. Leukemia. 2007 May;21(5):868-76. Epub 2007 Mar 15.

     2Leif W. Ellisen, Nadia Carlesso, Tao Cheng, David T. Scadden, and Daniel A. Haber  The Wilms tumor suppressor WT1 directs stage-specific quiescence and differentiation of human hematopoietic progenitor cells. EMBO J. 2001 Apr 17; 20(8): 1897–1909.

     3Liu-Yin, J. et al. (2008) Predictive value of minimal residual disease (MRD) monitoring by RQ-PCR in WT1 positive patients entered in the UK MRC AML-15 Trial. Blood 112, 259.

     4GabertJ, Beillard E  et al. Standardization  and quality control studies of real-time quantitative  reverse transcriptase polymerase chain reaction of fusion gene transcripts for residual disease detection in leukemia – a Europe Against Cancer program. Leukemia. 2003 Dec;17(12):2318-57.

     5van der Velden, V.H., Hochhaus, A., Cazzaniga, G., Szczepanski, T., Gabert, J., and van Dongen, J.J. (2003) Detection of minimal residual disease in hematologic malignancies by real-time quantitative PCR: principles, approaches, and laboratory aspects. Leukemia 17, 1013.

     6Beillard, E. et al. (2003) Evaluation of candidate control genes for diagnosis and residual disease detection in leukemic patients using ‘real- time’ quantitative reverse-transcriptase polymerase chain reaction (RQ- PCR) - a Europe against cancer program. Leukemia 17, 2474.

Ordering information:

• WT1 RQ Kit, 24 tests                                                                         Cat.No RQ-17-24

• WT1 RQ Kit, 48 tests                                                                        Cat.No RQ-17-48

Additiional reagents:

TriZ Reagent Kit                                                                                  Cat.No TRK-100

ReverZyme Kit                                                                                    Cat.No RT-1

Price: on request 

Reagents are for research use only (RUO)