[One-step multiplex RT-PCR for identifying common fusion transcripts in childhood acute lymphoblastic leukemia]
Xiao-Wen Chen1, Fei-Qiu Wen, Rong-Yu Lv
1Institute of Pediatrics, Shenzhen Children's Hospital, Shenzhen, Guangdong 518038, China. fwen62@126.com.
Objective:
To evaluate the efficiency of one-step multiplex RT-PCR for identifying four common fusion transcripts (TEL/AML1, E2A/PBX1, MLL/AF4 and BCR/ABL) in children with acute lymphoblastic leukemia (ALL).
Methods:
Total RNA was extracted from bone marrow samples of 76 children who were newly diagnosed with ALL between January 2003 and December 2010. These RNAs were analyzed for TEL/AML1, E2A/PBX1, MLL/AF4 and BCR/ABL by one-step multiplex RT-PCR or common nested-multiplex PCR. The PCR products were confirmed by DNA sequencing.
Results:
TEL/AML1 was found in 12 cases (the length of products was 298 bp in 9 cases and 259 bp in 3 cases), E2A/PBX1 was found in 3 cases (the length of products was 373 bp), BCR/ABL was found in 1 case (the length of products was 2 124 bp), and MLL/AF4 was found in 7 cases (the length of products was 427 bp in 1 case and 673 bp in 6 cases) using one-step multiplex RT-PCR combined with DNA sequencing. The results were consistent with those using common nested-multiplex PCR.
Conclusions:
One-step multiplex RT-PCR may be another alternative for detection of common fusion transcripts in children with ALL.
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