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Simplified strategies for minimal residual disease detection in B‐cell precursor acute lymphoblastic leukaemia
Author(s) -
LandmanParker J.,
Aubin J.,
Delabesse E.,
Tabone MD.,
Adam M.,
Millien C.,
Leboeuf D.,
BuzynVeil A.,
Dollfus C.,
Leverger G.,
Macintyre E. A.
Publication year - 1996
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.1996.d01-1917.x
Subject(s) - minimal residual disease , ethidium bromide , primer (cosmetics) , polymerase chain reaction , serial dilution , microbiology and biotechnology , acute lymphocytic leukemia , allele , calla , real time polymerase chain reaction , biology , medicine , bone marrow , immunology , gastroenterology , dna , pathology , antibody , genetics , lymphoblastic leukemia , gene , leukemia , chemistry , alternative medicine , organic chemistry , monoclonal antibody
We have developed a simplified fluorescent run‐off (FluRO) based IgH PCR strategy in order to facilitate follow‐up of large numbers of B‐cell precursor (BCP) acute lymphoblastic leukaemias (ALL) in a routine molecular diagnostic laboratory. DNA samples from 26 BCP‐ALL and one B‐cell line were amplified using IgH FR1 and FR2 consensus primers and analysed in parallel either by ethidium bromide non‐denaturing PAGE or, after rendering the PCR products fluorescent with an internal JH consensus primer, by high‐resolution analysis on an automated fragment analyser. The latter led to a minimum of one log increase in sensitivity of detection in 62% of alleles from 19 samples (16/28 in FR1; 11/15 in FR2) tested in parallel on log DNA dilutions, and to at least a 10 −2 level of sensitivity of detection in 15/19. The improved resolution allowed an approximate 20% increase in the number of clonal alleles detected, and consequently doubled the incidence of oligoclonality (6/26; 23%). Using these strategies, 6/17 (35%) of children analysed prospectively showed residual IgH positivity in the post induction complete remission bone marrow sample. Both early deaths occurred within this subgroup of patients and of the three of four surviving patients tested, two remained positive 2–3 months later. Although this simplified strategy is, as expected, less sensitive than anti‐V‐D‐J junction specific strategies, it enables detection of a category of ‘slow‐remitters’ which may have prognostic significance at a stage where therapeutic decisions are taken.