Establishment and application of a novel method for detecting MLL fusion genes of acute leukemia

This study was aimed to establish an efficient method to detect 10 common MLL fusion genes in patients with acute leukemia.Firstly,the relevant references and databases were searched to throughly inivestigate all fusion breakpoints;the primers and probes were designed according to nearly all the involved fusion types of gene.Then the multiplex real-time PCR system was established and optimized by using the established 16 positive plasmids and negative cell lines.Finally,the detection system was clinically evaluated by means of collected 54 samples of leukemia.The results indicated that the established detection system could efficiently detect all positive plasmids with sensibility to 10 copies.Four kinds of fusion gene types such as MLL-AF4,MLL-AF9,MLL-AF10,MLL-ELL could be detected in 54 samples,the sequencing of positive samples showed consistency of sequencing results with detection results.It is concluded that a novel multiplex real-time PCR detection method is established which can detect 10 common MLL fusion genes covering about 90% of the cases harboring MLL fusions.This method is fast,sensitive,specific and reliable,and should be an useful clinical tool for identification and management of leukemia patients with MLL fusions.