Application of BIOMED-2 Primers in Fixed and Decalcified Bone Marrow Biopsies
作者： Silke Lassmann , Ulrike V. Gerlach , Katja Technau-Ihling , Martin Werner , Paul Fisch
DOI: 10.1016/S1525-1578(10)60591-0
关键词: DNA extraction 、 Biopsy 、 Antibody 、 Follicular lymphoma 、 Bone marrow 、 Polymerase chain reaction 、 Bone decalcification 、 Biology 、 Molecular biology 、 B cell 、 Pathology and Forensic Medicine 、 Molecular medicine
摘要: The detection of clonality in lymphomas was recently improved by the BIOMED-2 approach, but analysis fixed tissues is limited. Here, we adapted protocol for examining immunoglobulin H (IgH) receptor rearrangements fixed, decalcified bone marrow biopsies (BMBs) B-cell non-Hodgkin's (B-NHL). study included 111 BMBs (12 formalin and 99 glutardialdehyde fixed), with B-NHL (n = 85), T-NHL 8), or reactive infiltrates 18). Initially, IgH FRIII polymerase chain reaction (PCR) crude DNA extracts from 75 glutardialdehyde-fixed (B-NHLs) using a standard seminested PCR resulted clonal peaks 46 (61.3%) compared 19 70 (27.1%) original protocol. Modifications to both extraction rate 26 36 (72.2%) primers, including 10 15 (66.7%) cases not detected our analysis. Moreover, introducing same modifications FRII region primers revealed (52.8%) B-NHLs 5 (7.1%) Together, regions modified increased 31 (86.1%), particularly histological evidence follicular lymphoma (FRIII, 70%; FRII, 90%). In summary, this provides an IgH-specific biopsies.
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