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Ancillary techniques in bone marrow pathology: molecular diagnostics on bone marrow trephine biopsies

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Abstract

Pathologic examination of trephine bone marrow (BM) biopsies plays a central role in the diagnosis and staging of haematological neoplasms and other disorders affecting haematopoiesis. Haematopathology has been profoundly influenced by the advent of molecular genetic techniques suitable for paraffin-embedded tissues, and certain applications, such as the determination of B- and T-cell clonality, belong to its standard diagnostic repertoire. Many of these molecular tests can be performed successfully with nucleic acids extracted from BM trephine biopsies, if some technical aspects specific to this template source such as various fixation and decalcification procedures are taken into consideration. The current indications for molecular BM diagnostics range from the confirmation of lymphoma involvement with gene rearrangement analysis, demonstration of tumor-specific translocations in lymphoid and chronic myeloproliferative disorders along to the detection of microorganisms or marrow involvement by soft tissue sarcomas. The availability of quantitative polymerase chain reaction techniques for the investigation of allelic imbalances and gene expression levels in paraffin-embedded material also open new avenues for research and advanced diagnostics. The molecular detection of minimal residual disease in haematological neoplasms, especially in the context of new treatment strategies, will provide future challenges. This article summarizes the current state of the art in molecular diagnostics applied to paraffin-embedded BM biopsies.

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References

  1. Akcan Y, Tuncer S, Hayran M, Sungur A, Unal S (1997) PCR on disseminated tuberculosis in bone marrow and liver biopsy specimens: correlation to histopathological and clinical diagnosis. Scand J Infect Dis 29:271–274

    Article  PubMed  CAS  Google Scholar 

  2. Arber DA (2000) Molecular diagnostic approach to non-Hodgkin's lymphoma. J Mol Diagn 2:178–190

    PubMed  CAS  Google Scholar 

  3. Aurer I, Juhbashi T, Sekine I, Tomonaga M, Gale RP (1993) Analysis of BCR/ABL abnormalities in mRNA from 20-year-old paraffin-embedded tissue for BCR/ABL rearrangement by polymerase chain reaction. Acta Haematol 90:5–7

    PubMed  CAS  Google Scholar 

  4. Bain BJ (2001) Bone marrow trephine biopsy. J Clin Pathol 54:737–742

    Article  PubMed  CAS  Google Scholar 

  5. Baxter EJ, Scott LM, Campbell PJ, East C, Fourouclas N, Swanton S, Vassiliou GS, Bench AJ, Boyd EM, Curtin N, Scott MA, Erber WN, Green AR (2005) Acquired mutation of the tyrosine kinase JAK2 in human myeloproliferative disorders. Lancet 365:1054–1061

    PubMed  CAS  Google Scholar 

  6. Bock O, Lehmann U, Kreipe H (2003) Quantitative intra-individual monitoring of BCR–ABL transcript levels in archival bone marrow trephines of patients with chronic myeloid leukemia. J Mol Diagn 5:54–60

    PubMed  CAS  Google Scholar 

  7. Bock O, Reising D, Kreipe H (2003) Multiplex RT-PCR for the detection of common BCR–ABL fusion transcripts in paraffin-embedded tissues from patients with chronic myeloid leukemia and acute lymphoblastic leukemia. Diagn Mol Pathol 12:119–123

    PubMed  CAS  Google Scholar 

  8. Braunschweig R, Baur AS, Delacretaz F, Bricod C, Benhattar J (2003) Contribution of IgH–PCR to the evaluation of B-cell lymphoma involvement in paraffin-embedded bone marrow biopsy specimens. Am J Clin Pathol 119:634–642

    Article  PubMed  CAS  Google Scholar 

  9. Brinckmann R, Kaufmann O, Reinartz B, Dietel M (2000) Specificity of PCR-based clonality analysis of immunoglobulin heavy chain gene rearrangements for the detection of bone marrow involvement by low-grade B-cell lymphomas. J Pathol 190:55–60

    Article  PubMed  CAS  Google Scholar 

  10. Browett PJ, Cooke HM, Secker-Walker LM, Norton JD (1989) Chromosome 22 breakpoints in variant Philadelphia translocations and Philadelphia-negative chronic myeloid leukemia. Cancer Genet Cytogenet 37:169–177

    Article  PubMed  CAS  Google Scholar 

  11. Bruggemann M, Pott C, Ritgen M, Kneba M (2004) Significance of minimal residual disease in lymphoid malignancies. Acta Haematol 112:111–119

    Article  PubMed  Google Scholar 

  12. Cabras AD, Kremer M, Schulz S, Werner M, Hummel M, Komminoth P, Höfler G, Höfler H (2000) Quality assessment in diagnostic molecular pathology: experience from a German–Austrian–Swiss multicenter trial. Virchows Arch 437:46

    Article  PubMed  CAS  Google Scholar 

  13. Cilloni D, Guerrasio A, Giugliano E, Scaravaglio P, Volpe G, Rege-Cambrin G, Saglio G (2002) From genes to therapy: the case of Philadelphia chromosome-positive leukemias. Ann N Y Acad Sci 963:306–312

    Article  PubMed  CAS  Google Scholar 

  14. Coad JE, Olson DJ, Christensen DR, Lander TA, Chibbar R, McGlennen RC, Brunning RD (1997) Correlation of PCR-detected clonal gene rearrangements with bone marrow morphology in patients with B-lineage lymphomas. Am J Surg Pathol 21:1047–1056

    Article  PubMed  CAS  Google Scholar 

  15. Cotelingam JD (2003) Bone marrow biopsy: interpretive guidelines for the surgical pathologist. Adv Anat Pathol 10:8–26

    PubMed  Google Scholar 

  16. Cross NC, Melo JV, Feng L, Goldman JM (1994) An optimized multiplex polymerase chain reaction (PCR) for detection of BCR–ABL fusion mRNAs in haematological disorders. Leukemia 8:186–189

    PubMed  CAS  Google Scholar 

  17. Darby AJ, Johnson PW (2002) Molecular remission and non-Hodgkin's lymphoma. Best Pract Res Clin Haematol 15:549–562

    Article  PubMed  CAS  Google Scholar 

  18. Deane M, McCarthy KP, Wiedemann LM, Norton JD (1991) An improved method for detection of B-lymphoid clonality by polymerase chain reaction. Leukemia 5:726–730

    PubMed  CAS  Google Scholar 

  19. Deininger MW, Goldman JM, Melo JV (2000) The molecular biology of chronic myeloid leukemia. Blood 96:3343–3356

    PubMed  CAS  Google Scholar 

  20. Derksen PW, Langerak AW, Kerkhof E, Wolvers-Tettero IL, Boor PP, Mulder AH, Vrints LW, Coebergh JW, van Krieken JH, Schuuring E, Kluin PM, van Dongen JJ (1999) Comparison of different polymerase chain reaction-based approaches for clonality assessment of immunoglobulin heavy-chain gene rearrangements in B-cell neoplasia. Mod Pathol 12:794–805

    PubMed  CAS  Google Scholar 

  21. Diebold J, Molina T, Camilleri-Broet S, Le Tourneau A, Audouin J (2000) Bone marrow manifestations of infections and systemic diseases observed in bone marrow trephine biopsy review. Histopathology 37:199–211

    Article  PubMed  CAS  Google Scholar 

  22. DiGiuseppe JA, Hartmann DP, Freter C, Cossman J, Mann RB (1997) Molecular detection of bone marrow involvement in intravascular lymphomatosis. Mod Pathol 10:33–37

    PubMed  CAS  Google Scholar 

  23. Dolken G (2001) Detection of minimal residual disease. Adv Cancer Res 82:133–185

    Article  PubMed  CAS  Google Scholar 

  24. Druker BJ, Sawyers CL, Kantarjian H, Resta DJ, Reese SF, Ford JM, Capdeville R, Talpaz M (2001) Activity of a specific inhibitor of the BCR–ABL tyrosine kinase in the blast crisis of chronic myeloid leukemia and acute lymphoblastic leukemia with the Philadelphia chromosome. N Engl J Med 344:1038–1042

    Article  PubMed  CAS  Google Scholar 

  25. Elenitoba-Johnson KS, Bohling SD, Mitchell RS, Brown MS, Robetorye RS (2000) PCR analysis of the immunoglobulin heavy chain gene in polyclonal processes can yield pseudoclonal bands as an artifact of low B cell number. J Mol Diagn 2:92–96

    PubMed  CAS  Google Scholar 

  26. Fend F, Weyrer K, Drach J, Schwaiger A, Umlauft F, Grunewald K (1993) Immunoglobulin gene rearrangement in plasma cell dyscrasias: detection of small clonal cell populations in peripheral blood and bone marrow. Leuk Lymphoma 10:223–229

    PubMed  CAS  Google Scholar 

  27. Fend F, Gschwendtner A, Gredler E, Thaler J, Dietze O (1994) Detection of monoclonal B-cell populations in decalcified, plastic-embedded bone marrow biopsies with the polymerase chain reaction. Am J Clin Pathol 102:850–855

    PubMed  CAS  Google Scholar 

  28. Fend F, Quintanilla-Martinez L, Kumar S, Beaty MW, Blum L, Sorbara L, Jaffe ES, Raffeld M (1999) Composite low grade B-cell lymphomas with two immunophenotypically distinct cell populations are true biclonal lymphomas. A molecular analysis using laser capture microdissection. Am J Pathol 154:1857–1866

    PubMed  CAS  Google Scholar 

  29. Fend F, Kremer M, Specht K, Quintanilla-Martinez L (2003) Laser microdissection in hematopathology. Pathol Res Pract 199:425–430

    Article  PubMed  CAS  Google Scholar 

  30. Gebhard S, Benhattar J, Bricod C, Meuge-Moraw C, Delacretaz F (2001) Polymerase chain reaction in the diagnosis of T-cell lymphoma in paraffin-embedded bone marrow biopsies: a comparative study. Histopathology 38:37–44

    Article  PubMed  CAS  Google Scholar 

  31. Gleissner B, Gokbuget N, Bartram CR, Janssen B, Rieder H, Janssen JW, Fonatsch C, Heyll A, Voliotis D, Beck J, Lipp T, Munzert G, Maurer J, Hoelzer D, Thiel E (2002) Leading prognostic relevance of the BCR–ABL translocation in adult acute B-lineage lymphoblastic leukemia: a prospective study of the German multicenter trial group and confirmed polymerase chain reaction analysis. Blood 99:1536–1543

    Article  PubMed  CAS  Google Scholar 

  32. Godfrey TE, Kim SH, Chavira M, Ruff DW, Warren RS, Gray JW, Jensen RH (2000) Quantitative mRNA expression analysis from formalin-fixed, paraffin-embedded tissues using 5′ nuclease quantitative reverse transcription–polymerase chain reaction. J Mol Diagn 2:84–91

    PubMed  CAS  Google Scholar 

  33. Gong JZ, Zheng S, Chiarle R, De Wolf-Peeters C, Palestro G, Frizzera G, Inghirami G (1999) Detection of immunoglobulin kappa light chain rearrangements by polymerase chain reaction. An improved method for detecting clonal B-cell lymphoproliferative disorders. Am J Pathol 155:355–363

    PubMed  CAS  Google Scholar 

  34. Grunewald K, Lyons J, Hansen-Hagge TE, Janssen JW, Feichtinger H, Bartram CR (1991) Molecular genetic analysis of DNA obtained from fixed, air dried or paraffin embedded sources. Ann Hematol 62:108–114

    Article  PubMed  CAS  Google Scholar 

  35. Hanamura A, Kinoshita T, Kurokawa T, Nagai H, Murate T, Nagasaka T, Mori H, Saito H (1999) Molecular evaluation of bone marrow involvement in peripheral T-cell lymphoma with a PCR-mediated RNase protection assay. Int J Hematol 70:283–289

    PubMed  CAS  Google Scholar 

  36. Hanson CA, Kurtin PJ, Katzmann JA, Hoyer JD, Li CY, Hodnefield JM, Meyers CH, Habermann TM, Witzig TE (1999) Immunophenotypic analysis of peripheral blood and bone marrow in the staging of B-cell malignant lymphoma. Blood 94:3889–3896

    Google Scholar 

  37. Heinmoller E, Renke B, Beyser K, Dietmaier W, Langner C, Ruschoff J (2001) Piffalls in diagnostic molecular pathology-significance of sampling error. Virchows Arch 439:504–511

    Article  PubMed  CAS  Google Scholar 

  38. Hoeve MA, Krol AD, Philippo K, Derksen PW, Veenendaal RA, Schuuring E, Kluin PM, van Krieken JH (2000) Limitations of clonality analysis of B cell proliferations using CDR3 polymerase chain reaction. Mol Pathol 53:194–200

    Article  PubMed  CAS  Google Scholar 

  39. James C, Ugo V, Le Couedic JP, Staerk J, Delhommeau F, Lacout C, Garcon L, Raslova H, Berger R, Bennaceur-Griscelli A, Villeval JL, Constantinescu SN, Casadevall N, Vainchenker W (2005) A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature 434:1144–1148

    Article  PubMed  CAS  Google Scholar 

  40. Koopmans M, Monroe SS, Coffield LM, Zaki SR (1993) Optimization of extraction and PCR amplification of RNA extracts from paraffin-embedded tissue in different fixatives. J Virol Methods 43:189–204

    Article  PubMed  CAS  Google Scholar 

  41. Krafft AE, Duncan BW, Bijwaard KE, Taubenberger JK, Lichy JH (1997) Optimization of the isolation and amplification of RNA from formalin-fixed, paraffin-embedded tissue: the Armed Forces Institute of Pathology experience and literature review. Mol Diagn 2:217–230

    Article  PubMed  CAS  Google Scholar 

  42. Kralovics R, Passamonti F, Buser AS, Teo SS, Tiedt R, Passweg JR, Tichelli A, Cazzola M, Skoda RC (2005) A gain-of-function mutation of JAK2 in myeloproliferative disorders. N Engl J Med 352:1779–1790

    Article  PubMed  CAS  Google Scholar 

  43. Kremer M, Cabras AD, Fend F, Schulz S, Schwarz K, Hoefler H, Werner M (2000) PCR analysis of IgH-gene rearrangements in small lymphoid infiltrates microdissected from sections of paraffin-embedded bone marrow biopsy specimens. Hum Pathol 31:847–853

    Article  PubMed  CAS  Google Scholar 

  44. Kremer M, Sandherr M, Geist B, Cabras AD, Hofler H, Fend F (2001) Epstein–Barr virus-negative Hodgkin's lymphoma after mycosis fungoides: molecular evidence for distinct clonal origin. Mod Pathol 14:91–97

    Article  PubMed  CAS  Google Scholar 

  45. Kremer M, Spitzer M, Mandl-Weber S, Stecker K, Schmidt B, Hofler H, Quintanilla-Martinez L, Fend F (2003) Discordant bone marrow involvement in diffuse large B-cell lymphoma: comparative molecular analysis reveals a heterogeneous group of disorders. Lab Invest 83:107–114

    PubMed  CAS  Google Scholar 

  46. Kreuzer KA, Lass U, Nagel S, Ellerbrok H, Pauli G, Pawlaczyk-Peter B, Siegert W, Huhn D, Schmidt CA (2000) Applicability of an absolute quantitative procedure to monitor intra-individual bcr/abl transcript kinetics in clinical samples from chronic myelogenous leukemia patients. Int J Cancer 86:741–746

    Article  PubMed  CAS  Google Scholar 

  47. Laurent E, Talpaz M, Kantarjian H, Kurzrock R (2001) The BCR gene and philadelphia chromosome-positive leukemogenesis. Cancer Res 61:2343–2355

    PubMed  CAS  Google Scholar 

  48. Lehmann U, Bock O, Langer F, Kreipe H (2001) Demonstration of light chain restricted clonal B-lymphoid infiltrates in archival bone marrow trephines by quantitative real-time polymerase chain reaction. Am J Pathol 159:2023–2029

    PubMed  CAS  Google Scholar 

  49. Lehmann U, Kreipe H (2001) Real-time PCR analysis of DNA and RNA extracted from formalin-fixed and paraffin-embedded biopsies. Methods 25:409–418

    Article  PubMed  CAS  Google Scholar 

  50. Levine RL, Wadleigh M, Cools J, Ebert BL, Wernig G, Huntly BJ, Boggon TJ, Wlodarska I, Clark JJ, Moore S, Adelsperger J, Koo S, Lee JC, Gabriel S, Mercher T, D'Andrea A, Frohling S, Dohner K, Marynen P, Vandenberghe P, Mesa RA, Tefferi A, Griffin JD, Eck MJ, Sellers WR, Meyerson M, Golub TR, Lee SJ, Gilliland DG (2005) Activating mutation in the tyrosine kinase JAK2 in polycythemia vera, essential thrombocythemia, and myeloid metaplasia with myelofibrosis. Cancer Cell 7:387–397

    Article  PubMed  CAS  Google Scholar 

  51. Lewis F, Maughan NJ, Smith V, Hillan K, Quirke P (2001) Unlocking the archive-gene expression in paraffin-embedded tissue. J Pathol 195:66–71

    Article  PubMed  CAS  Google Scholar 

  52. Limpens J, de Jong D, van Krieken JH, Price CG, Young BD, van Ommen GJ, Kluin PM (1991) Bcl-2/JH rearrangements in benign lymphoid tissues with follicular hyperplasia. Oncogene 6:2271–2276

    PubMed  CAS  Google Scholar 

  53. Limpens J, Stad R, Vos C, de Vlaam C, de Jong D, van Ommen GJ, Schuuring E, Kluin PM (1995) Lymphoma-associated translocation t(14;18) in blood B cells of normal individuals. Blood 85:2528–2536

    PubMed  CAS  Google Scholar 

  54. Markoulatos P, Siafakas N, Moncany M (2002) Multiplex polymerase chain reaction: a practical approach. J Clin Lab Anal 16:47–51

    Article  PubMed  CAS  Google Scholar 

  55. Masuda N, Ohnishi T, Kawamoto S, Monden M, Okubo K (1999) Analysis of chemical modification of RNA from formalin-fixed samples and optimization of molecular biology applications for such samples. Nucleic Acids Res 27:4436–4443

    Article  PubMed  CAS  Google Scholar 

  56. McCarthy KP, Sloane JP, Wiedemann LM (1990) Rapid method for distinguishing clonal from polyclonal B cell populations in surgical biopsy specimens. J Clin Pathol 43:429–432

    Article  PubMed  CAS  Google Scholar 

  57. McCarthy KP, Sloane JP, Kabarowski JH, Matutes E, Wiedemann LM (1992) A simplified method of detection of clonal rearrangements of the T-cell receptor-gamma chain gene. Diagn Mol Pathol 1:173–179

    Article  PubMed  CAS  Google Scholar 

  58. Miranda RN, Mark HF, Medeiros LJ (1994) Fluorescent in situ hybridization in routinely processed bone marrow aspirate clot and core biopsy sections. Am J Pathol 145:1309–1314

    PubMed  CAS  Google Scholar 

  59. Nagasaka T, Lai R, Chen YY, Chen W, Arber DA, Chang KL, Weiss LM (2000) The use of archival bone marrow specimens in detecting B-cell non-Hodgkin's lymphomas using polymerase chain reaction methods. Leuk Lymphoma 36:347–352

    PubMed  CAS  Google Scholar 

  60. Olavarria E, Kanfer E, Szydlo R, Kaeda J, Rezvani K, Cwynarski K, Pocock C, Dazzi F, Craddock C, Apperley JF, Cross NC, Goldman JM (2001) Early detection of BCR–ABL transcripts by quantitative reverse transcriptase-polymerase chain reaction predicts outcome after allogeneic stem cell transplantation for chronic myeloid leukemia. Blood 97:1560–1565

    Article  PubMed  CAS  Google Scholar 

  61. Pajor L, Lacza A, Kereskai L, Jakso P, Egyed M, Ivanyi JL, Radvanyi G, Dombi P, Pal K, Losonczy H (2004) Increased incidence of monoclonal B-cell infiltrate in chronic myeloproliferative disorders. Mod Pathol 12:1521–1530

    Article  Google Scholar 

  62. Peterson LA, Brunning RD (2001) Bone marrow specimen processing. In: Knowles DM (ed) Neoplastic hematopathology. Lippincott Williams & Wilkins, Philadelphia, pp 1391–1406

    Google Scholar 

  63. Pittaluga S, Tierens A, Dodoo YL, Delabie J, De Wolf-Peeters C (1999) How reliable is histologic examination of bone marrow trephine biopsy specimens for the staging of non-Hodgkin lymphoma? A study of hairy cell leukemia and mantle cell lymphoma involvement of the bone marrow trephine specimen by histologic, immunohistochemical, and polymerase chain reaction techniques. Am J Clin Pathol 111:179–184

    PubMed  CAS  Google Scholar 

  64. Provan AB, Hodges E, Smith AG, Smith JL (1992) Use of paraffin wax embedded bone marrow trephine biopsy specimens as a source of archival DNA. J Clin Pathol 45:763–765

    Article  PubMed  CAS  Google Scholar 

  65. Radich JP, Gooley T, Bryant E, Chauncey T, Clift R, Beppu L, Edmands S, Flowers ME, Kerkof K, Nelson R, Appelbaum FR (2001) The significance of bcr–abl molecular detection in chronic myeloid leukemia patients “late,” 18 months or more after transplantation. Blood 98:1701–1707

    Article  PubMed  CAS  Google Scholar 

  66. Ravandi F, Cortes J, Albitar M, Arlinghaus R, Qiang Guo J, Talpaz M, Kantarjian HM (1999) Chronic myelogenous leukaemia with p185(BCR/ABL) expression: characteristics and clinical significance. Br J Haematol 107:581–586

    Article  PubMed  CAS  Google Scholar 

  67. Salto-Tellez M, Shelat SG, Benoit B, Rennert H, Carroll M, Leonard DG, Nowell P, Bagg A (2003) Multiplex RT-PCR for the detection of leukemia-associated translocations: validation and application to routine molecular diagnostic practice. J Mol Diagn 5:231–236

    PubMed  CAS  Google Scholar 

  68. Schmid C, Isaacson PG (1992) Bone marrow trephine biopsy in lymphoproliferative disease. J Clin Pathol 45:745–750

    Article  PubMed  CAS  Google Scholar 

  69. Schulz S, Cabras AD, Kremer M, Weirich G, Miethke T, Bosmuller HC, Hofler H, Werner M, Fend F (2005) Species identification of mycobacteria in paraffin-embedded tissues: frequent detection of nontuberculous mycobacteria. Mod Pathol 18(2):274–282

    Article  PubMed  CAS  Google Scholar 

  70. Sotlar K, Escribano L, Landt O, Mohrle S, Herrero S, Torrelo A, Lass U, Horny HP, Bultmann B (2003) One-step detection of c-kit point mutations using peptide nucleic acid-mediated polymerase chain reaction clamping and hybridization probes. Am J Pathol 162:737–746

    PubMed  CAS  Google Scholar 

  71. Specht K, Kremer M, Muller U, Dirnhofer S, Rosemann M, Hofler H, Quintanilla-Martinez L, Fend F (2002) Identification of cyclin D1 mRNA overexpression in B-cell neoplasias by real-time reverse transcription-PCR of microdissected paraffin sections. Clin Cancer Res 8:2902–2911

    PubMed  CAS  Google Scholar 

  72. Specht K, Haralambieva E, Bink K, Kremer M, Mandl-Weber S, Koch I, Tomer R, Hofler H, Schuuring E, Kluin PM, Fend F, Quintanilla-Martinez L (2004) Different mechanisms of cyclin D1 overexpression in multiple myeloma revealed by fluorescence in situ hybridization and quantitative analysis of mRNA levels. Blood 104:1120–1126

    Article  PubMed  CAS  Google Scholar 

  73. Specht K, Richter T, Müller U, Walch A, Werner M, Höfler H (2001) Quantitative gene expression analysis in microdissected archival formalin-fixed, paraffin-embedded tumor tissue. Am J Pathol 158:419–429

    PubMed  CAS  Google Scholar 

  74. Srinivasan M, Sedmak D, Jewell S (2002) Effect of fixatives and tissue processing on the content and integrity of nucleic acids. Am J Pathol 161:1961–1971

    PubMed  CAS  Google Scholar 

  75. Stacchini A, Demurtas A, Godio L, Martini G, Antinoro V, Palestro G (2003) Flow cytometry in the bone marrow staging of mature B-cell neoplasms. Cytometry B Clin Cytom 54:10–18

    Article  PubMed  CAS  Google Scholar 

  76. Taylor ML, Sehgal D, Raffeld M, Obiakor H, Akin C, Mage RG, Metcalfe DD (2004) Demonstration that mast cells, T cells, and B cells bearing the activating kit mutation D816V occur in clusters within the marrow of patients with mastocytosis. J Mol Diagn 6:335–342

    PubMed  CAS  Google Scholar 

  77. Tbakhi A, Totos G, Pettay JD, Myles J, Tubbs RR (1999) The effect of fixation on detection of B-cell clonality by polymerase chain reaction. Mod Pathol 12:272–278

    PubMed  CAS  Google Scholar 

  78. Thiele J, Zirbes TK, Kvasnicka HM, Fischer R (1999) Focal lymphoid aggregates (nodules) in bone marrow biopsies: differentiation between benign hyperplasia and malignant lymphoma—a practical guideline. J Clin Pathol 52:294–300

    PubMed  CAS  Google Scholar 

  79. van Dongen JJ, Langerak AW, Bruggemann M, Evans PA, Hummel M, Lavender FL, Delabesse E, Davi F, Schuuring E, Garcia-Sanz R, van Krieken JH, Droese J, Gonzalez D, Bastard C, White HE, Spaargaren M, Gonzalez M, Parreira A, Smith JL, Morgan GJ, Kneba M, Macintyre EA (2003) Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: report of the BIOMED-2 concerted action BMH4-CT98-3936. Leukemia 17:2257–2317

    Article  PubMed  Google Scholar 

  80. Viswanatha D, Foucar K (2003) Hodgkin and non-Hodgkin lymphoma involving bone marrow. Semin Diagn Pathol 20:196–210

    Article  PubMed  Google Scholar 

  81. Weirich G, Funk A, Hoepner I, Heider U, Noll S, Putz B, Fellbaum C, Hofler H (1995) PCR-based assays for the detection of monoclonality in non-Hodgkin's lymphoma: application to formalin-fixed, paraffin-embedded tissue and decalcified bone marrow samples. J Mol Med 73:235–241

    Article  PubMed  CAS  Google Scholar 

  82. Weirich G, Sandherr M, Fellbaum C, Richter T, Schmidt L, Kinjerski T, Dietzfelbinger H, Rastetter J, Hofler H (1998) Molecular evidence of bone marrow involvement in advanced case of Tgammadelta lymphoma with secondary myelofibrosis. Hum Pathol 29:761–765

    Article  PubMed  CAS  Google Scholar 

  83. Wickham CL, Boyce M, Joyner MV, Sarsfield P, Wilkins BS, Jones DB, Ellard S (2000) Amplification of PCR products in excess of 600 base pairs using DNA extracted from decalcified, paraffin wax embedded bone marrow trephine biopsies. Mol Pathol 53:19–23

    Article  PubMed  CAS  Google Scholar 

  84. Wilson GA, Vandenberghe EA, Pollitt RC, Rees DC, Goodeve AC, Peake IR, Reilly JT (2000) Are aberrant BCR–ABL transcripts more common than previously thought? Br J Haematol 111:1109–1111

    Article  PubMed  CAS  Google Scholar 

  85. Zhou XG, Sandvej K, Gregersen N, Hamilton-Dutoit SJ (1999) Detection of clonal B cells in microdissected reactive lymphoproliferations: possible diagnostic pitfalls in PCR analysis of immunoglobulin heavy chain gene rearrangement. Mol Pathol 52:104–110

    PubMed  CAS  Google Scholar 

  86. Zoubek A, Ladenstein R, Windhager R, Amann G, Fischmeister G, Kager L, Jugovic D, Ambros PF, Gadner H, Kovar H (1998) Predictive potential of testing for bone marrow involvement in Ewing tumor patients by RT-PCR: a preliminary evaluation. Int J Cancer 79:56–60

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

We thank Mrs. Birgit Geist for her excellent technical support. This study was supported in part by the Deutsche Forschungsgemeinschaft (DFG), grant FE 597/1 to FF, LQ-M and MK.

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  1. Institute of Pathology, Technical University Munich, Ismaningerstrasse 22, 81675, Munich, Germany

    Falko Fend, Markus Kremer & Katja Specht

  2. Institute of Pathology, Hannover Medical School, Hannover, Germany

    Oliver Bock

  3. Institute of Pathology, GSF Research Center for Environment and Health, Neuherberg, Oberschleissheim, Germany

    Leticia Quintanilla-Martinez

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  1. Falko Fend
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Correspondence to Falko Fend.

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Fend, F., Bock, O., Kremer, M. et al. Ancillary techniques in bone marrow pathology: molecular diagnostics on bone marrow trephine biopsies. Virchows Arch 447, 909–919 (2005). https://doi.org/10.1007/s00428-005-0069-1

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