Detection Limit of Monoclonal B-Cells Using Multiplex PCR and Laser-Induced Fluorescence Capillary Electrophoresis
이성학, 문연숙, Song Byung-Hoo, 이형남, 이아원, 정은선, 최영진, 이교영, 강창석, 박경신,
소속 상세정보
이성학 ( Lee Sung-Hak )
Catholic University Seoul St. Mary’s Hospital Department of Hospital Pathology
문연숙 ( Moon Yeon-Sook )
Inha University School of Medicine Department of Laboratory Medicine
( Song Byung-Hoo )
Catholic University Seoul St. Mary’s Hospital Department of Hospital Pathology
이형남 ( Lee Hyung-Nam )
Catholic University Seoul St. Mary’s Hospital Department of Hospital Pathology
이아원 ( Lee Ah-Won )
Catholic University Seoul St. Mary’s Hospital Department of Hospital Pathology
정은선 ( Jung Eun-Sun )
Catholic University College of Medicine Department of Hospital Pathology
최영진 ( Choi Yeong-Jin )
Catholic University College of Medicine Department of Hospital Pathology
이교영 ( Lee Kyo-Young )
Catholic University College of Medicine Department of Hospital Pathology
강창석 ( Kang Chang-Suk )
Catholic University Seoul St. Mary’s Hospital Department of Hospital Pathology
박경신 ( Park Gyeong-Sin )
Catholic University Seoul St. Mary’s Hospital Department of Hospital Pathology
KMID : 0357920110450060582
Abstract
Background : The identification of monoclonality has been widely used for making diagnoses of lymphoproliferative lesions. Awareness of the sensitivity and detection limit of the technique used would be important for the data to be convincing.
Methods : We investigated the minimum requirement of cells and sensitivity of gel electrophoresis (GE) and laser-induced fluorescence capillary electrophoresis (LFCE) for identifying IgH gene rearrangement using BIOMED-2 protocols. DNA extracted from Raji cells were diluted serially with peripheral blood mononuclear cells (PBMNCs) DNA. DNA from mixtures of diffuse large B-cell lymphoma (DLBCL) and reactive lymph nodes were also serially diluted.
Results : For Raji cells, the detection limit was 62 and 16 cell-equivalents for GE and LFCE, respectively. In the condition with PBMNCs mixture, 2.5% and 1.25% of clonal cells was the minimum requirement for GE and LFCE, respectively. In 23% of DLBCL cells in tissue section, the detection limit was 120 and 12 cell-equivalents for GE and LFCE, respectively. In 3.2% of DLBCL cells, that was 1,200 and 120 cell-equivalents for GE and LFCE, respectively.
Conclusions : These results show that LFCE method is more sensitive than GE and the sensitivity of clonality detection can be influenced by the amount of admixed normal lymphoid cells.
키워드
Electrophoresis; capillary; Monoclonality; Lymphoproliferative disorders; BIOMED-2
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