JOURNAL BROWSE
Search
Advanced SearchSearch Tips
Multiplex RT-PCR Assay for Detection of Common Fusion Transcripts in Acute Lymphoblastic Leukemia and Chronic Myeloid Leukemia Cases
facebook(new window)  Pirnt(new window) E-mail(new window) Excel Download
 Title & Authors
Multiplex RT-PCR Assay for Detection of Common Fusion Transcripts in Acute Lymphoblastic Leukemia and Chronic Myeloid Leukemia Cases
Limsuwanachot, Nittaya; Siriboonpiputtana, Teerapong; Karntisawiwat, Kanlaya; Chareonsirisuthigul, Takol; Chuncharunee, Suporn; Rerkamnuaychoke, Budsaba;
  PDF(new window)
 Abstract
Background: Acute lymphoblastic leukemia (ALL) is a heterogeneous disease which requires a risk-stratified approach for appropriate treatment. Specific chromosomal translocations within leukemic blasts are important prognostic factors that allow identification of relevant subgroups. In this study, we developed a multiplex RT-PCR assay for detection of the 4 most frequent translocations in ALL (BCR-ABL, TEL-AML1, MLL-AF4, and E2A-PBX1). Materials and Methods: A total of 214 diagnosed ALL samples from both adult and pediatric ALL and 14 cases of CML patients (154 bone marrow and 74 peripheral blood samples) were assessed for specific chromosomal translocations by cytogenetic and multiplex RT-PCR assays. Results: The results showed that 46 cases of ALL and CML (20.2%) contained the fusion transcripts. Within the positive ALL patients, the most prevalent cryptic translocation observed was mBCR-ABL (p190) at 8.41%. In addition, other genetic rearrangements detected by the multiplex PCR were 4.21% TEL-AML1 and 2.34% E2A-PBX1, whereas MLL-AF4 exhibited negative results in all tested samples. Moreover, MBCR-ABL was detected in all 14 CML samples. In 16 samples of normal karyotype ALL (n
 Keywords
Acute lymphoblastic leukemia;chronic myeloid leukemia;multiplex RT-PCR;risk stratification;
 Language
English
 Cited by
 References
1.
Awan T, Iqbal Z, Aleem A, et al (2012). Five most common prognostically important fusion oncogenes are detected in the majority of pakistani pediatric acute lymphoblastic leukemia patients and are strongly associated with disease biology and treatment outcome. Asian Pac J Cancer Prev, 13, 5469-75. crossref(new window)

2.
Bhatia P, Binota J, Varma N, et al (2012). Incidence of common chimeric fusion transcripts in B-cell acute lymphoblastic leukemia: an Indian perspective. Acta Haematol, 129, 17-9.

3.
Borowitz MJ, Hunger SP, Carroll AJ, et al (1993). Predictability of the t(1;19)(q23;p13) from surface antigen phenotype: implications for screening cases of childhood acute lymphoblastic leukemia for molecular analysis: a Pediatric Oncology Group study. Blood, 82, 1086-91.

4.
Branford S, Rudzki Z, Walsh S, et al (2003). Detection of BCRABL mutations in patients with CML treated with imatinib is virtually always accompanied by clinical resistance, and mutations in the ATP phosphate-binding loop (P-loop) are associated with a poor prognosis. Blood, 102, 276-83. crossref(new window)

5.
Colleoni GW, Jhanwar SC, Ladanyi M, Chen B (2000). Comparison of a multiplex reverse transcriptasepolymerase chain reaction for BCR-ABL to fluorescence in situ hybridization, Southern blotting, and conventional cytogenetics in the monitoring of patients with Ph1-positive leukemias. Diagn Mol Pathol, 9, 203-9. crossref(new window)

6.
Crist WM, Carroll AJ, Shuster JJ, et al (1990). Poor prognosis of children with pre-B acute lymphoblastic leukemia is associated with the t(1;19)(q23;p13): a Pediatric Oncology Group study. Blood, 76, 117-22.

7.
Dunlap J, Beadling C, Warrick A, et al (2012). Multiplex high-throughput gene mutation analysis in acute myeloid leukemia. Hum Pathol, 43, 2167-76. crossref(new window)

8.
Elia L, Mancini M, Moleti L, et al (2003). A multiplex reverse transcriptase-polymerase chain reaction strategy for the diagnostic molecular screening of chimeric genes: a clinical evaluation on 170 patients with acute lymphoblastic leukemia. Haematologica, 88, 275-9.

9.
Elias MH, Baba AA, Azlan H, et al (2014). BCR-ABL kinase domain mutations, including 2 novel mutations in imatinib resistant Malaysian chronic myeloid leukemia patients-Frequency and clinical outcome. Leuk Res, 38, 454-9. crossref(new window)

10.
Elmaagacli AH (2007). Molecular methods used for detection of minimal residual disease following hematopoietic stem cell transplantation in myeloid disorders. Methods Mol Biol, 1109, 187-207.

11.
Erba HP (2015). Molecular monitoring to improve outcomes in patients with chronic myeloid leukemia in chronic phase: importance of achieving treatment-free remission. Am J Hematol, 90, 242-9. crossref(new window)

12.
Gabert J, Beillard E, van der Velden VH, et al (2003). Standardization and quality control studies of ‘real-time' quantitative reverse transcriptase polymerase chain reaction of fusion gene transcripts for residual disease detection in leukemia - a Europe Against Cancer program. Leukemia, 17, 2318-57. crossref(new window)

13.
Goldman J (2005). Monitoring minimal residual disease in BCRABL-positive chronic myeloid leukemia in the imatinib era. Curr Opin Hematol, 12, 33-9. crossref(new window)

14.
Greaves MF, Maia AT, Wiemels JL, Ford AM (2003). Leukemia in twins: lessons in natural history. Blood, 102, 2321-33. crossref(new window)

15.
Greaves MF, Wiemels J (2003). Origins of chromosome translocations in childhood leukaemia. Nat Rev Cancer, 3, 639-49. crossref(new window)

16.
Harrison CJ (2000). The management of patients with leukaemia:the role of cytogenetics in this molecular era. Br J Haematol, 108, 19-30. crossref(new window)

17.
Hughes T, Saglio G, Branford S, et al (2009). Impact of baseline BCR-ABL mutations on response to nilotinib in patients with chronic myeloid leukemia in chronic phase. J Clin Oncol, 27, 4204-10. crossref(new window)

18.
Kadhom AE, Abdul-Majeed BA, AL-Hadad SA, Day P (2015). Expression of TEL-AML1 and E2A-PBX1 by real-time PCR in pediatric ALL in Baghdad. Int J Adv Res, 3, 820-6.

19.
Kim SH, Kim D, Kim DW, et al (2009). Analysis of Bcr-Abl kinase domain mutations in Korean chronic myeloid leukaemia patients: poor clinical outcome of P-loop and T315I mutation is disease phase dependent. Hematol Oncol, 27, 190-7. crossref(new window)

20.
Krauter J, Peter W, Pascheberg U, et al (1998). Detection of karyotypic aberrations in acute myeloblastic leukaemia: a prospective comparison between PCR/FISH and standard cytogenetics in 140 patients with de novo AML. Br J Haematol, 103, 72-8. crossref(new window)

21.
Krivtsov AV, Armstrong SA (2007). MLL translocations, histone modifications and leukaemia stem-cell development. Nat Rev Cancer, 7, 823-33. crossref(new window)

22.
Liang DC, Shih LY, Yang CP, et al (2002). Multiplex RTPCR assay for the detection of major fusion transcripts in Taiwanese children with B-lineage acute lymphoblastic leukemia. Med Pediatr Oncol, 39, 12-7. crossref(new window)

23.
Look AT (1997). Oncogenic transcription factors in the human acute leukemias. Science, 278, 1059-64. crossref(new window)

24.
Mazloumi SHM, Madhumathi DS, Appaji L, Prasannakumari P (2012). Combined study of cytogenetics and fluorescence in situ hybridization (FISH) analysis in childhood acute lymphoblastic leukemia (ALL) in a Tertiary Cancer Centre in South India. Asian Pac J Cancer Prev, 13, 3825-7. crossref(new window)

25.
Mellentin JD, Nourse J, Hunger SP, Smith SD, Cleary ML (1990). Molecular analysis of the t(1;19) breakpoint cluster region in pre-B cell acute lymphoblastic leukemias. Genes Chromosomes Cancer, 2, 239-47. crossref(new window)

26.
Muntean AG, Hess JL (2012). The pathogenesis of mixed-lineage leukemia. Annu Rev Pathol, 7, 283-301. crossref(new window)

27.
Park TS, Lee ST, Song J, et al (2008). MLL rearrangement with t(6;11)(q15;q23) as a sole abnormality in a patient with de novo acute myeloid leukemia: conventional cytogenetics, FISH, and multicolor FISH analyses for detection of rare MLL-related chromosome abnormalities. Cancer Genet Cytogenet, 187, 50-3. crossref(new window)

28.
Piwkham D, Siriboonpiputtana T, Beuten J, et al (2015). Mutation screening and association study of the folylpolyglutamate synthetase (FPGS) gene with susceptibility to childhood acute lymphoblastic leukemia. Asian Pac J Cancer Prev, 16, 4727-32. crossref(new window)

29.
Raanani P, Ben-Bassat I, Gan S, et al (2004). Assessment of the response to imatinib in chronic myeloid leukemia patients--comparison between the FISH, multiplex and RT-PCR methods. Eur J Haematol, 73, 243-50. crossref(new window)

30.
Salto-Tellez M, Shelat SG, Benoit B, et al (2003). Multiplex RTPCR for the detection of leukemia-associated translocations: validation and application to routine molecular diagnostic practice. J Mol Diagn, 5, 231-6. crossref(new window)

31.
Sarriera JE, Albitar M, Estrov Z, et al (2001). Comparison of outcome in acute myelogenous leukemia patients with translocation (8;21) found by standard cytogenetic analysis and patients with AML1/ETO fusion transcript found only by PCR testing. Leukemia, 15, 57-61. crossref(new window)

32.
Scurto P, Hsu RM, Kane JR, et al (1998). A multiplex RT-PCR assay for the detection of chimeric transcripts encoded by the risk-stratifying translocations of pediatric acute lymphoblastic leukemia. Leukemia, 12, 1994-2005. crossref(new window)

33.
Shaffer JG, McGowan-Jordan J, Schmid M (2013). Neoplasia. In 'ISCN 2013: An International System for Human Cytogenetic Nomenclature'. S Karger AG, Inc., Basel pp 88-96.

34.
Shaikh MS, Adil SN, Shaikh MU, Khurshid M (2014). Frequency of chromosomal abnormalities in Pakistani adults with acute lymphoblastic leukemia. Asian Pac J Cancer Prev, 15, 9495-8. crossref(new window)

35.
Sudhakar N, Rajalekshmy KR, Rajkumar T, Nancy KN (2011). RT-PCR and real-time PCR analysis of E2A-PBX1, TELAML1, mBCR-ABL and MLL-AF4 fusion gene transcripts in de novo B-lineage acute lymphoblastic leukaemia patients in south India. J Genet, 90, 349-53. crossref(new window)

36.
Tahira B, Asif M, Khan S, et al (2015). Detection of BCR/ABL fusion gene by hematological and cytogenetical analysis in chronic myeloid leukemia patients in Quetta, Pakistan. Asian Pac J Cancer Prev, 16, 3793-7. crossref(new window)

37.
Testoni N, Marzocchi G, Luatti S, et al (2009). Chronic myeloid leukemia: a prospective comparison of interphase fluorescence in situ hybridization and chromosome bandinganalysis for the definition of complete cytogenetic response: a study of the GIMEMA CML WP. Blood, 114, 4939-43. crossref(new window)

38.
van Dongen JJ, Macintyre EA, Gabert JA, et al (1999). Standardized RT-PCR analysis of fusion gene transcripts from chromosome aberrations in acute leukemia for detection of minimal residual disease. Report of the BIOMED-1 Concerted Action: investigation of minimal residual disease in acute leukemia. Leukemia, 13, 1901-28. crossref(new window)

39.
Vardiman JW, Thiele J, Arber DA, et al (2009). The 2008 revision of the World Health Organization (WHO) classification of myeloid neoplasms and acute leukemia: rationale and important changes. Blood, 114, 937-51. crossref(new window)

40.
Von Goessel H, Jacobs U, Semper S, et al (2009). Cluster analysis of genomic ETV6-RUNX1 (TEL-AML1) fusion sites in childhood acute lymphoblastic leukemia. Leuk Res, 33, 1082-8. crossref(new window)

41.
Wiemels JL, Greaves M (1999). Structure and possible mechanisms of TEL-AML1 gene fusions in childhood acute lymphoblastic leukemia. Cancer Res, 59, 4075-82.

42.
Wiemels JL, Leonard BC, Wang Y, et al (2002). Site-specific translocation and evidence of postnatal origin of the t(1;19) E2A-PBX1 fusion in childhood acute lymphoblastic leukemia. Proc Natl Acad Sci U S A, 99, 15101-6. crossref(new window)

43.
Wongboonma W, Thongnoppakhun W, Auewarakul CU (2011). BCR-ABL kinase domain mutations in tyrosine kinase inhibitors-naïve and -exposed Southeast Asian chronic myeloid leukemia patients. Exp Mol Pathol, 92, 259-65.

44.
Xiong FF, Li BS, Zhang CX, et al (2013). A pipeline with multiplex reverse transcription polymerase chain reaction and microarray for screening of chromosomal translocations in leukemia. Biomed Res Int.

45.
Yang YL, Lin SR, Chen JS, et al (2010). Multiplex reverse transcription-polymerase chain reaction as diagnostic molecular screening of 4 common fusion chimeric genes in Taiwanese children with acute lymphoblastic leukemia. J Pediatr Hematol Oncol, 32, 323-30. crossref(new window)

46.
Ye YX, Zhou J, Zhou YH, et al (2014). Clinical significance of BCR-ABL fusion gene subtypes in chronic myelogenous and acute lymphoblastic leukemias. Asian Pac J Cancer Prev, 15, 9961-6. crossref(new window)

47.
Zafar I (2014). Molecular Genetic Studies on 167 Pediatric all patients from different areas of pakistan confirm a low frequency of the favorable prognosis fusion oncogene TELAML1 (t 12; 21) in underdeveloped countries of the region. Asian Pac J Cancer Prev, 15, 3541-6. crossref(new window)

48.
Zelent A, Greaves M, Enver T (2004). Role of the TEL-AML1 fusion gene in the molecular pathogenesis of childhood acute lymphoblastic leukaemia. Oncogene, 23, 4275-83. crossref(new window)