Improvement of clonality detection rate in multiple myeloma using fluorescent IgH PCR with different sets of primers

Citation
V. Welterlin et al., Improvement of clonality detection rate in multiple myeloma using fluorescent IgH PCR with different sets of primers, J HEMATH ST, 9(6), 2000, pp. 983-991
Citations number
35
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Hematology,"Medical Research Diagnosis & Treatment
Journal title
JOURNAL OF HEMATOTHERAPY & STEM CELL RESEARCH
ISSN journal
1525-8165 → ACNP
Volume
9
Issue
6
Year of publication
2000
Pages
983 - 991
Database
ISI
SICI code
1525-8165(200012)9:6<983:IOCDRI>2.0.ZU;2-S
Abstract
The IgH rearrangement provides a useful marker of clonality in B-cell malig nancies and amplification of this rearrangement is the method of choice to monitor the residual tumor cells in multiple myeloma (MM). The critical poi nt of this analysis was the false-negative rate observed at diagnosis in pa tients presenting tumor cells well above the limit of detection. The aim of this study was therefore to increase the clonality detection rate by IgH p olymerase chain reaction (PCR). Bone marrow DNA from 37 MM patients were an alyzed at diagnosis. IgH PCR with agarose gel detection was performed betwe en framework regions FR3 and FR1, both in combination with 5 different prim ers in FR4. Fluorescent IgH PCR with highly resolutive capillary electropho resis was used to improve the detection and to size clonal PCR products. Si xty-two percent of the clonal rearrangements were initially detected with J HD primer specific to the JH segments 1,2,4,5. The use of JH3 and JH6 homol ogous primers increased the detection rate to 78%, whereas a consensus JH p rimer only reached 67% of positivity. The lowest detection rates were obtai ned with JHExt and JH3 with a detection of respectively 43 and 14%. However , three rearrangements were exclusively amplified by JHExt and two addition al cases were detected by JH3. The combined use of primers yielded the best score with 89% of positivity. With Genescan analysis, two additional cases showed a monoclonal rearrangement improving the detection rate to 95%. The use of multiple sets of primers along with a highly sensitive genescan ana lysis makes possible the follow-up of minimal residual disease for most MM patients.