Abstract: Objective：To establish and evaluate a real-time fluorescence quantitative PCR (qRT-PCR) method for detecting human glycogen synthase kinase 3β ( GSK3β ) mRNA and its splice variant GSK3βΔex8+9.fragment. Methods：The target fragments of GSK3β and GSK3βΔex8+9 were amplified from the total RNA extracted from the peripheral blood or bone marrow of the patients with chronic myelocytic leukemia (CML) or with the blast crisis of CML, Then, their plasmid standard preparation and standard curve were constructed to establish a qRT-PCR method, and its reproducibility and specificity were evaluated. Results：The GSK3β (133 bp) and GSK3βΔex8+9 (219 bp) fragments were amplified and their corresponding qRT-PCR methods were established successfully. The standard curve showed a fine linear relationship between the cycle threshold (CT) and the template concentration, and the coefficients of correlation (r) were 0.997 and 0.999, respectively. Both of qRT-PCR methods had high reproducibility, with the intra-assay and inter-assay coefficients of variation (CV) less than 2%. The specificity analysis demonstrated that only 133 bp of target fragment could be amplified from the patients′ mRNA by the qRT-PCR method for detecting GSK3β but not GSK3βΔex8+9. Conclusion：The established methods have fine reproducibility and specificity, which can be used to quantitatively detect the mRNA levels of GSK3β and GSK3βΔex8+9.