Rapid isolation of translocation breakpoints in chronic myeloid and acute promyelocytic leukaemia

Summary Isolation and sequencing of the translocation breakpoint in chronic myeloid leukaemia‐(CML) and acute promyelocytic leukaemia (APML) may help to elucidate the mechanism of translocation and provide a molecular marker for monitoring of minimal residual disease. Amplification across the transl...

Full description

Saved in:

Bibliographic Details
Published in:	British journal of haematology Vol. 149; no. 2; pp. 231 - 236
Main Authors:	Bartley, Paul A., Martin‐Harris, Michael H., Budgen, Bradley J., Ross, David M., Morley, Alexander A.
Format:	Journal Article
Language:	English
Published:	Oxford, UK Blackwell Publishing Ltd 01-04-2010 Blackwell
Subjects:	acute promyelocytic leukaemia BCR‐ABL1 Biological and medical sciences Chromosome aberrations chronic myeloid leukaemia Fusion Proteins, bcr-abl - genetics Hematologic and hematopoietic diseases Humans Leukemia, Myelogenous, Chronic, BCR-ABL Positive - genetics Leukemia, Promyelocytic, Acute - genetics Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis Medical genetics Medical sciences PML‐RARA polymerase chain reaction Polymerase Chain Reaction - methods Translocation, Genetic translocations Chromosomal aberration Acute myelogenous leukemia acute promyelocytic leukaemia Myeloproliferative syndrome PML-RARA Hybrid gene Chromosome translocation Promyelocytic leukemia Breakpoint Hematology Acute Chronic myelogenous leukemia Malignant hemopathy Abl1 gene chronic myeloid leukaemia Polymerase chain reaction bcr gene Abnormal chromosome E17 Abnormal chromosome D15 BCR-ABL1 Cytogenetics translocations Abnormal chromosome Molecular biology Abnormal chromosome C11 Chronic promyelocytic leukemia Cancer M3 acute myelocytic leukemia
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	Summary Isolation and sequencing of the translocation breakpoint in chronic myeloid leukaemia‐(CML) and acute promyelocytic leukaemia (APML) may help to elucidate the mechanism of translocation and provide a molecular marker for monitoring of minimal residual disease. Amplification across the translocation breakpoint was performed in samples from 91 patients with CML and 15 patients with APML using single‐tube multiplex polymerase chain reaction (PCR) involving 308 primers for CML and 40 primers for APML. Nonspecific amplification was removed by a modification of PCR, termed sequential hybrid primer PCR (SHP‐PCR), which involved two sequential rounds of PCR, each of which used a low concentration of a specially designed hybrid primer. The resultant amplified material was sequenced. The method as finally developed was simple quick and robust. The translocation breakpoint was successfully isolated and sequenced in all 106 samples. The strategy of highly multiplexed PCR followed by SHP‐PCR is thus an effective method for isolating the translocation breakpoint in CML and APML. It may also be applicable to other haematological disorders characterised by translocation, deletion or inversion.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0007-1048 1365-2141
DOI:	10.1111/j.1365-2141.2009.08071.x