Detection of Clonal Immunoglobulin and T-Cell Receptor Gene Rearrangements in Childhood Acute Lymphoblastic Leukemia Using a Low-Cost PCR Strategy

Author(s)

Assumpcao, Juliana Godoy ^[1] ; Ganazza, Monica Aparecida ^[1] ; de Araujo, Marcela ^[1] ; Silva, Ariosto Siqueira ^[2] ; Scrideli, Carlos Alberto ^[3] ; Brandalise, Silvia Regina ^{[4, 1]} ; Yunes, Jose Andres ^[1]

Affiliation

^[1] Ctr Infantil Boldrini, Mol Biol Lab, BR-13083210 Campinas, SP - Brazil

^[2] H Lee Moffitt Canc Ctr & Res Inst, Dept Integrat Math Oncol, Tampa, FL - USA

^[3] Univ Sao Paulo, Dept Pediat, Ribeirao Preto Med Sch, BR-14049 Ribeirao Preto - Brazil

^[4] Univ Estadual Campinas, Dept Pediat, Campinas, SP - Brazil

• Document type: Journal article
• Source: PEDIATRIC BLOOD & CANCER; v. 55, n. 7, p. 1278-1286, DEC 15 2010.
• Web of Science Citations: 2

Abstract

Background Imunoglobulin (Ig) and T cell receptor (TCR) gene rearrangements function as specific markers for minimal residual disease (MRD) which is one of the best predictors of outcome in childhood acute lymphoblastic leukemia (ALL) We recently reported on the prognostic value of MRD during the induction of remission through a simplified PCR method Here we report on gene rearrangement frequencies and offer guidelines for the application of the technique Procedure Two hundred thirty three children had DNA extracted from bone marrow Ig and TCR gene rearrangements were amplified using consensus primers and conventional PCR PCR products were submitted to homo/heteroduplex analysis A computer program was designed to define combinations of targets for clonal detection using a minimum set of primers and reactions Results At least one clonal marker could be detected in 98% of the patients and two markers in approximately 80% The most commonly rear ringed genes in precursor B cell ALL were IgH (75%) TCRD (59%) IgK (55%), and TCRG (54%) The most commonly rearranged genes for TALL were TCRG (100%) and TCRD (24%) The sensitivity of primers was limited to the detection of 1 leukemic cell among 100 normal cells Conclusions We propose that eight PCR reactions per ALL subtype would allow for the detection of two markers in most cases In addition these reactions ire suitable for MRD monitoring especially when aiming the selection of patients with high MRD levels (>= 10(-2)) at the end of induction therapy Such an approach would be very useful in centers with limited financial resources Pediatr Blood Cancer 2010 55 1278-1286 (C) 2010 Wiley Liss Inc (AU)

• FAPESP's process: 05/02279-6 - Minimal residual disease study in acute lymphoblastic leukemia of children and teenager
• Grantee: Carlos Alberto Scrideli
• Support Opportunities: Regular Research Grants