Fluorescence quantitative PCR detection of WT_1 gene expression in leukemias

Objctive:To establish a fluorescense quantitative RT-PCR(FQ-RT-PCR)method for detection of WT 1 gene expression.To elucidate the expression of WT 1 in all types of leukemias.Methods:The conventional RT-PCR was used to amplify WT 1 gene and β-actin gene from K562 cells.After gel extraction,purification and quantification by a photometer,quantitative stand template was constructed.Using FQ-RT-PCR,we measured the peripheral blood or bone marrow samples from 80 leukemia patients and 10 normal voluteer′s peripheral blood samples.Results:The expression of WT 1 gene in all types of leukemias was significantly higher than that in normal controls.WT 1 expression levels was significantly higher in myeloid than in lymphoid.Patients with M 5 expressed significantly lower levels.Conclusion:The established FQ-RT-PCR method is sensitive and specific.The expression of WT 1 gene was relatively high in all types of leukemias compared with normal peripheral blood cells.