Quantitative assessment of contaminating tumor cells in autologous peripheral blood stem cells of B-cell non-Hodgkin lymphomas using immunoglobulin heavy chain gene allele-specific oligonucleotide real-time quantitative-polymerase chain reaction.
A real-time quantitative-polymerase chain reaction (RQ-PCR) targeting the immunoglobulin heavy chain (IgH) gene has been used for the quantification of minimal residual disease (MRD) in B-cell hematological malignancies. In non-Hodgkin lymphoma (NHL), experimental costs are increased, as a large number of primer-probe sets are required because of diversity, due to somatic and ongoing mutations of the IgH gene. We developed an allele-specific oligonucleotide (ASO) combined with a germline consensus probe-based RQ-PCR assay and examined MRD in peripheral blood stem cells (PBSC). The IgH consensus probes were adapted in seven (50%) of 14 amplifiable cases. Patients with heavily contaminating tumor cells in PBSC relapsed after PBSC transplantation. Our strategy will contribute to the development of a cost-efficient, precisely quantitative and systemic detection assay for MRD in NHL.
Yashima
Akiko
A
Department of Pathology, Iwate Medical University School of Medicine, Uchimaru 19-1, 020-8505 Morioka, Japan. ayashima@iwate-med.ac.jp
Maesawa
Chihaya
C
Uchiyama
Michihiro
M
Tarusawa
Mitsu
M
Satoh
Takashi
T
Satoh
Mamiko
M
Enomoto
Sanae
S
Sugawara
Ken
K
Numaoka
Hideharu
H
Murai
Kazunori
K
Utsugisawa
Taiju
T
Ishida
Yoji
Y
Masuda
Tomoyuki
T
D000293:Adolescent; D000328:Adult; D000368:Aged; D000483:Alleles; D001483:Base Sequence; D016384:Consensus Sequence; D017931:DNA Primers / Q000191:economics / Q000592:standards; D005260:Female; D005803:Genes, Immunoglobulin / Q000235:genetics*; D006801:Humans; D016393:Lymphoma, B-Cell / Q000628:therapy*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D018365:Neoplasm, Residual / Q000175:diagnosis*; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D009841:Oligonucleotides; D036102:Peripheral Blood Stem Cell Transplantation / Q000379:methods* / Q000592:standards; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods* / Q000592:standards; D012680:Sensitivity and Specificity; D014182:Transplantation, Autologous
D017931:DNA Primers; D009841:Oligonucleotides