Development of immunoglobulin variable heavy chain gene consensus probes with conjugated 3′ minor groove binder groups for monitoring minimal residual disease in childhood acute lymphoblastic leukaemia
- M Uchiyama1,
- C Maesawa1,
- A Yashima1,
- T Itabashi1,
- T Satoh1,
- M Tarusawa2,
- M Endo2,
- Y Takahashi4,
- S Sasaki4,
- S Tsuchiya5,
- Y Ishida3,
- T Masuda1
- 1Department of Pathology, Iwate Medical University School of Medicine, Uchimaru 19-1, Morioka 020-8505, Japan
- 2Department of Paediatrics, Iwate Medical University School of Medicine
- 3Division of Haematology, Third Department of Internal Medicine, Iwate Medical University School of Medicine
- 4Department of Paediatrics, Hirosaki University School of Medicine, 036-8562 Hirosaki, Japan
- 5Department of Paediatric Oncology, Research Institute of Development, Aging and Cancer, Tohoku University, 980-0872 Sendai, Japan
- Correspondence to: Assistant Professor C Maesawa Department of Pathology, Iwate Medical University School of Medicine, Uchimaru 19-1, Morioka 020-8505, Japan; chihayaiwate-med.ac.jp
- Accepted 4 June 2003
Abstract
Aims: To develop immunoglobulin heavy chain variable (VH) gene probes that are shorter and more flexible in position for monitoring minimal residual disease (MRD) in childhood leukaemia (ALL), using minor groove binder (MGB) technology.
Methods: All VH germline sequences registered in the database were aligned and the consensus regions were determined. The reliability of the MGB probes was compared with non-MGB probes in all 24 cases of ALL.
Results: Ten MGB probes (16 to 18 mers) were designed that enabled all the germline sequences on the database to be analysed, whereas the conventional non-MGB probes (21 to 27 mers) did not allow the analysis of four of the VH1 and five of the VH3 germline sequences. The sequencing results in five of the 24 cases of ALL were not matched to the non-MGB probes.
Conclusions: MGB technology allows shorter probes to be designed, enabling MRD to be detected in childhood ALL. This would provide a considerable reduction in cost for a large MRD study.
- IgH monoclonality
- minor groove binder probe
- allele specific oligonucleotide real time quantitative polymerase chain reaction
- ASO RQ-PCR, allele specific oligonucleotide real time quantitative polymerase chain reaction
- ALL, acute lymphoblastic leukaemia
- IgH, immunoglobulin heavy chain
- MRD, minimal residual disease
- MGB, minor groove binder
- Tm, melting temperature
- VH, IgH variable region
