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Simple multiplex RT-PCR for identifying common fusion transcripts in childhood acute leukemia.
Int J Lab Hematol. 2008 Aug; 30(4):286-91.IJ

Abstract

Nonrandom gene rearrangements have been demonstrated in leukemic cells at diagnosis. These genetic abnormalities are associated with specific types, clinical characteristics, and prognosis of acute leukemia. Common fusion transcripts in childhood acute lymphoblastic leukemia (ALL) are TEL-AML1, E2A-PBX, MLL-AF4, and BCR-ABL (p190) and in acute nonlymphoblastic leukemia (ANLL) are AML-ETO, PML-RARA, and CBFB-MYH11. Reverse transcription-polymerase chain reaction (RT-PCR) for detection of each individual fusion transcript is impractical and time consuming. The purpose of this study was to develop simple RT-PCR methods to identify common fusion transcripts of newly diagnosed acute leukemia in children. Total RNA was extracted from bone marrow samples of children diagnosed with acute leukemia. Multiplex RT-PCR panel A (ALL) included primers for TEL-AML1, E2A-PBX, MLL-AF4, and BCR-ABL (p190) whereas panel B (ANLL) composed of primers for AML-ETO, PML-RARA, and CBFB-MYH11. Known leukemic cell lines were used to serve as positive controls. Eighty three children diagnosed with ALL (n = 63) and ANLL (n = 20) were included in this study. Fusion transcripts could be identified using multiplex RT-PCR panel A for ALL and panel B for ANLL in 26/83 (31.3%) cases. In ALL samples, we found TEL-AML1 = 16/63 (25.4%), E2A-PBX = 3/63 (4.8%), MLL-AF4 = 1/63 (1.6%), and BCR-ABL = 1/63 (1.6%). Four cases of AML1-ETO (20%) and one PML-RARA (5%) were found in ANLL samples. In conclusion, our simple multiplex RT-PCR for detection of fusion transcripts in childhood acute leukemia was found to be a rapid, accurate, and effective method.

Authors+Show Affiliations

Department of Pediatrics and Research Center, Faculty of Medicine, Ramathibodi Hospital, Bangkok, Thailand. rasam@mahidol.ac.thNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18665825

Citation

Pakakasama, S, et al. "Simple Multiplex RT-PCR for Identifying Common Fusion Transcripts in Childhood Acute Leukemia." International Journal of Laboratory Hematology, vol. 30, no. 4, 2008, pp. 286-91.
Pakakasama S, Kajanachumpol S, Kanjanapongkul S, et al. Simple multiplex RT-PCR for identifying common fusion transcripts in childhood acute leukemia. Int J Lab Hematol. 2008;30(4):286-91.
Pakakasama, S., Kajanachumpol, S., Kanjanapongkul, S., Sirachainan, N., Meekaewkunchorn, A., Ningsanond, V., & Hongeng, S. (2008). Simple multiplex RT-PCR for identifying common fusion transcripts in childhood acute leukemia. International Journal of Laboratory Hematology, 30(4), 286-91. https://doi.org/10.1111/j.1751-553X.2007.00954.x
Pakakasama S, et al. Simple Multiplex RT-PCR for Identifying Common Fusion Transcripts in Childhood Acute Leukemia. Int J Lab Hematol. 2008;30(4):286-91. PubMed PMID: 18665825.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Simple multiplex RT-PCR for identifying common fusion transcripts in childhood acute leukemia. AU - Pakakasama,S, AU - Kajanachumpol,S, AU - Kanjanapongkul,S, AU - Sirachainan,N, AU - Meekaewkunchorn,A, AU - Ningsanond,V, AU - Hongeng,S, PY - 2008/7/31/pubmed PY - 2008/10/10/medline PY - 2008/7/31/entrez SP - 286 EP - 91 JF - International journal of laboratory hematology JO - Int J Lab Hematol VL - 30 IS - 4 N2 - Nonrandom gene rearrangements have been demonstrated in leukemic cells at diagnosis. These genetic abnormalities are associated with specific types, clinical characteristics, and prognosis of acute leukemia. Common fusion transcripts in childhood acute lymphoblastic leukemia (ALL) are TEL-AML1, E2A-PBX, MLL-AF4, and BCR-ABL (p190) and in acute nonlymphoblastic leukemia (ANLL) are AML-ETO, PML-RARA, and CBFB-MYH11. Reverse transcription-polymerase chain reaction (RT-PCR) for detection of each individual fusion transcript is impractical and time consuming. The purpose of this study was to develop simple RT-PCR methods to identify common fusion transcripts of newly diagnosed acute leukemia in children. Total RNA was extracted from bone marrow samples of children diagnosed with acute leukemia. Multiplex RT-PCR panel A (ALL) included primers for TEL-AML1, E2A-PBX, MLL-AF4, and BCR-ABL (p190) whereas panel B (ANLL) composed of primers for AML-ETO, PML-RARA, and CBFB-MYH11. Known leukemic cell lines were used to serve as positive controls. Eighty three children diagnosed with ALL (n = 63) and ANLL (n = 20) were included in this study. Fusion transcripts could be identified using multiplex RT-PCR panel A for ALL and panel B for ANLL in 26/83 (31.3%) cases. In ALL samples, we found TEL-AML1 = 16/63 (25.4%), E2A-PBX = 3/63 (4.8%), MLL-AF4 = 1/63 (1.6%), and BCR-ABL = 1/63 (1.6%). Four cases of AML1-ETO (20%) and one PML-RARA (5%) were found in ANLL samples. In conclusion, our simple multiplex RT-PCR for detection of fusion transcripts in childhood acute leukemia was found to be a rapid, accurate, and effective method. SN - 1751-5521 UR - https://www.unboundmedicine.com/medline/citation/18665825/Simple_multiplex_RT_PCR_for_identifying_common_fusion_transcripts_in_childhood_acute_leukemia_ L2 - https://doi.org/10.1111/j.1751-553X.2007.00954.x DB - PRIME DP - Unbound Medicine ER -