Revista Română de Medicină de Laborator Vol. 20, Nr. 1/4, Martie 2012 Multiplex PCR approach for detection of fusion genes in acute lymphoblastic and myeloid leukemia PCR multiplex pentru detectia genelor de fuziune in leucemia acuta mieloida si limfoblastica Dumitru Jardan 1* , Rodica Talmaci 2 , Cerasela Jardan 1 , Daniel Coriu 2 , Anca Colita 2 , Andrei Colita 2 , Constantin Arion 1 1. Carol Davila University of Medicine and Pharmacy, Department of Pediatrics Clinical Institute Fundeni, Bucharest, Romania. 2. Carol Davila University of Medicine and Pharmacy, Department of Hematology Clinical Institute Fundeni, Bucharest, Romania. Abstract Acute leukemias are a group of heterogeneous diseases with respect to clinical features and acquired genetic aberrations. Since the discovery of BCR-ABL fusion gene a lot of other fusion genes were and are con- tinuing to be discovered in leukemia patients. Many of these fusion genes are believed to be pathognomonic to disease and can be used as prognostic factors for the evolution of disease. In this study we performed develop- ment and optimization of a multiplex PCR assay for detection of 8 most common fusion genes in AML and ALL, namely: PML-RARα, AML1-ETO, CBFβ-MYH11, BCR-ABL, MLL-AF4, E2A-PBX1, SIL-TAL and TEL- AML1.Results: Our multiplex PCR assay allowed multiplexing of primers for detection of 8 most common fusion genes in acute leukemia. This allowed risk stratification for 36.5% of the patients. Furthermore identification of these fusion genes allowed MRD monitoring using nested PCR or real-time PCR for quantitative evaluation of MRD. Overall a very good performance was obtained with no non-specific amplifications and 100% correlation with commercial HemaVision kits. In comparison with HemaVision kits this assay allows identification of the most frequent 8 fusion genes at a considerably lower price per patient. Keywords: Fusion genes, multiplex RT-PCR, acute leukemia Rezumat Leucemiile acute sunt un grup de boli heterogene in ceea ce priveste evolutia clinica si anomaliile genetice prezentate. De la descoperirea genei de fuziune BCR-ABL1 multe alte gene de fuziune au fost si sunt in continuare descrise la pacientii cu leucemie.O parte din aceste gene de fuziune sunt considerate patognomonice bolii si pot fi folosite ca factori de prognostic. In acests tudiu am dezvoltat si optimizat o metoda PCR multiplex pentru detectia a 8 gene de fuziune cu frecventa cea mai mare in LAM si LAL: PML-RARα, AML1-ETO, CBFβ-MYH11, BCR-ABL, MLL-AF4, E2A-PBX1, SIL-TAL si TEL-AML1. Rezultate: Metoda noastra, PCR multiplex, a permis combinarea * Corresponding author:Dumitru Jardan, Departament of Hematology, Fundeni Clinical Institute, Sos. Fundeni 258, Sector 2, Bucharest, Romania. Tel.: 0735300999, Email: d.jardan@gmail.com 15 Original article