iCMLf Forum 2014 - Dr Pilleux (Chile) - Implementation of qPCR

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Information about iCMLf Forum 2014 - Dr Pilleux (Chile) - Implementation of qPCR
Science-Technology

Published on March 10, 2014

Author: jangeissler

Source: authorstream.com

Implementation of Quantitative PCR of p210 BCR-ABL fusion transcript for Chronic Myeloid Leukemia patients at Valdivia Hospital, Chile. : Implementation of Quantitative PCR of p210 BCR-ABL fusion transcript for Chronic Myeloid Leukemia patients at Valdivia Hospital, Chile. Dr. Lilian Pilleux Valdivia Hospital Universidad Austral de Chile Valdivia, Chile PowerPoint Presentation: CHILE Valdivia The aim of our project is to monitor molecular response converted to the international scale of CML patients from southern Chile. : The aim of our project is to monitor molecular response converted to the international scale of CML patients from southern Chile. Chile : Chile Population 2013 : 17.556.815 inhabitants Hospital of Valdivia (Population CML) Región de Los Ríos 382.741 Región de Los Lagos 867.315 Región de Aysén 107.915 Región de Magallanes _ 160.164 1.518.135 Native ancestry / rurality Hematology specialists 5 Adult Hematologists 4 Children Hematooncologists Valdivia: Hospital of Valdivia: 28 patients in control 14 patients died last decade 2013 → access to: Cytogenetics FISH Valdivia Conventional PCR Quantitative PCR Mutations Study Santiago Valdivia CML Study and Therapy Time Line at Valdivia: 1988 National Program Antineoplastic Drugs 2002 Cualitative PCR - Valdivia 2002 Ministry of Health - Imatinib 10 Patients 2010 Ministry of Health Finances TKI 2004 Imatinib GIPAP 2006 Cytogenetics Lab - Santiago 2010 RT-QPCR BCR/ABL-Santiago 2008 Cytogenetics Lab - Valdivia 2011 RT-QPCR BCR/ABL-Valdivia 2013 CML Study and Therapy Time Line at Valdivia Access to Imatinib , Nilotinib and Dasatinib . Multistep Proyect: Multistep Proyect STEP 1 1999 Investigation Project: STEP 1 1999 Investigation Project Hematology Unit , Faculty of Medicine Universidad Austral de Chile ( UACh ). “ Using DNA probes marked with radioisotopes for the control, diagnosis and evaluation of malignant and infectious pathologies" F unded by: International Atomic Energy Agency (IAEA) Chilean Commission Nuclear Energy UACh PowerPoint Presentation: Provided the equipment of our molecular biology laboratory . + Hospital → Floor Space Qualitative PCR study for hemato -oncological diseases (children and adults) particularly acute leukemias and CML. PowerPoint Presentation:  It was necessary to gather a number of samples for weeks or months before performing the tests → inconvenient for clinical decisions. 2000-2006 STEP 2 2007-2010: STEP 2 2007-2010 Ensure the sustainability of the laboratory over time. COLDAS Project (Teaching – Health Assistance Agreement) “ Creating a regional referral center for the diagnosis of hemato -oncological diseases and viral hepatitis (2007-2010)” Standardization of the most prevalent genetic alterations in acute leukemias , chronic myeloproliferative disorders and lymphoma. Initiation of quantitative PCR for t(9;22) p210 Purchase new equipment. Biochemist position Studies for genetic alterations implemented 2007-2012 : Studies for genetic alterations implemented 2007-2012 LEUKEMIAS BCR-ABL p190 t(9;22) (q34;q11) ALL, CML BCR-ABL p210 t(9;22) (q34;q11) ALL, CML MLL-AF4 t(4;11) (q21;q23) ALL E2A-PBX1 t(1;19) (q23;p13) ALL TEL-AML1 t(12;21) (p13;q22). ALL PML-RARA t(15;17) (q22;q21) APL CBFB-MYH11 inv16 (p13;q22) AML AML1-ETO t(8;21) (q22;q22) AML SIL-TAL1 disruption at 1p32 T-ALL  LYMPHOMAS t(2;5) B-Cell Diffuse Large Cell Lymphoma t(11;14) Mantle Cell Lymphoma t(14;18) Folicular Lymphoma t(3;14) B-Cell Diffuse Large Cell Lymphoma t(11,18) MALT Lymphoma  OTHER DISEASES Mutación V617F de JAK2 PV, MF N- myc . Neuroblastoma (ALL: Acute Lymphoblastic Leukemia; AML: Acute Myeloid Leukemia; PV: policitemia vera; MF: Myelofibrosis ) STEP 3 2010 -2012: STEP 3 2010 -2012 The biochemist position was funded by the hospital. NO FUNDING for the determinations itself. We appealed to Pharmaceutical laboratories to finance the fee for quantitative PCR p210 BCR-ABL determinations performed at our lab for CML patients treated with TKI. Novartis - Bristoll Myers Squibb Allowed the continuity of molecular biology determinations. In addition permitted the purchase of some new equipment. STEP 4 2012-2013: STEP 4 2012-2013 The molecular biology laboratory is assumed as a hospital responsibility, for both technical and administrative matters, inserting itself in the Pathology Department. Valid equipment provided in the previous stages was maintained and the remaining equipment was renewed through funds provided by the Ministry of Health of Chile. 2013: 2013 Implementation MMR monitoring with IVD/CE reagents including a calibrator with an assigned value according to the “International Genetic Reference Panel for the quantitation of BCR-ABL translocation by RQ-PCR”. Tracking curves with the number of copies normalized and converted to the International Scale . Key Learnings: Key Learnings Education Investigation Outreach Health Care Next Steps: Next Steps Ask Ministry of Health to name Valdivia Hospital as reference for RT-Q-PCR in CML patients for public hospitals of southern Chile (3 - 4 fold ). B egin m utation analisis by DNA s equencing buying reagents with iCMLf grant while using university equipment . Conclusions: Conclusions CML therapy is in Progress in Chile Chile (Dr. Soledad Undurraga ) 405 patients (2001-2012) 329 (81%) cytogenetics 109 (27%) molecular monitoring 37 (9%) ≥ MMR L aboratory is an essential tool in clinical practice to eventually achieve cure of CML. Valdivia Hospital Response Patients (n) PHR* 3 CHR 6 CCyR 5 MMR 14 Total 28 *(1 T315I mutation ; 1 discontinuation ; 1 Failure )

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