The clonal determination of B-cell lymphoproliferative diseases by immunoglobulin heavy chain (IgH) rearrangement by polymerase chain reaction (PCR) is widely used. In this study we report our experiences with B cell clonality analysis using multiplex PCR amplification followed by heteroduplex analysis, which was found to be essential for the efficient resolution of monoclonal bands within polyclonal backgrounds, in formalin-fixed paraffinembedded tissue.
1. Diss Tc, Pan L. Polymerase chain reaction in the assessment of lymphomas. Cancer Surv 1997; 30:21-44.
2. Davis Re, Warnke Ra, Dorfman Rf. Utility of molecular genetic analysis for the diagnosis of neoplasia in morphologically and immunophenotypically equivocal hematolymphoid lesions. Cancer 1991; 67:2890-2899.
3. Beishuizen A, Verhoeven Ma, Mol Ej, Breit Tm, Wolvers-Tettero Il, Van Dongen Jj. Detection of immunoglobulin heavy-chain gene rearrangements by Southern blot analysis: recommendations for optimal results. Leukemia 1993; 7:2045-2053.
4. Tonegawa S. Somatic generation of antibody diversity. Nature 1983; 302, 575-581.
5. Van Dongen Jj, Langerak Aw, Bruggemann M, Evans Pa, Hummel M, et al. 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 2003; 17(12):2257-2317.
6. Diss Tc, Pan L, Peng H. Sources of DNA for detecting B cell monoclonality using PCR. J Clin Pathol 1994; 47:493-496.
7. Wan Jh, Trainor Kj, Brisco Mj. Monoclonality in B cell lymphoma detected in paraffin wax embedded sections using the polymerase chain reaction. J Clin Pathol 1990; 43:888-890.
8. Gong Jz, Zheng S, Chiarle R, De Wolf-Peeters C, Palestro G, et al. Detection of immunoglobulin kappa light chain rearrangements by polymerase chain reaction. An improved method for detecting clonal B-cell lymphoproliferative disorders. Am J Pathol. 1999; 155(2):355-363.
9. Weiss, S.W. (1994) WHO: Histological typing of soft tissue tumors. Berlin, Springer, 1994.
10. Elaine S. J. The 2008 WHO classification of lymphomas: implications for clinical practice and translational research Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, 2008.
11. Coelho Siqueira Sa, Ferreira Alves Va, Beitler B, Otta Mm, Nascimento Saldiva Ph. Contribution of immunohistochemistry to small B-cell lymphoma classification. Appl Immunohistochem Mol Morphol. 2006;14(1):1-6.
12. Salles G, De Jong D, Xie W, Rosenwald A, Chhanabhai M, et al. Prognostic significance of immunohistochemical biomarkers in diffuse large B-cell lymphoma: a study from the Lunenburg Lymphoma Biomarker Consortium. Blood 2011; 117(26):7070-7078.
13. Trainor Kj, Brisco Mj, Wan Jh, Neoh S, Grist S, et al. Gene rearrangement in B- and T-lymphoproliferative disease detected by the polymerase chain reaction. Blood. 1991; 78(1):192-196.
14. Kneba M, Bolz I, Linke B, Hiddemann W. Analysis of rearranged T-cell receptor -chain genes by polymerase chain reaction (PCR) DNA sequencing and automated high resolution PCR fragment analysis. Blood 1995; 86:3930-3937.
15. Garcia-Sanz R, Lopez-Perez R, Langerak Aw, Gonzalez D, Chillon Mc, et al. Heteroduplex PCR analysis of rearranged immunoglobulin genes for clonality assessment in multiple myeloma. Haematologica. 1999;84(4):328-335.
16. Alaibac M, Filotico R, Giannella C, Paradiso A, Labriola A, et al. The effect of fixation type on DNA extracted from paraffin-embedded tissue for PCR studies in dermatopathology. Dermatology 1997; 195: 105-107.
17. An Sf, Fleming Ka. Removal of inhibitor(s) of the polymerase chain reaction from formalin fixed, paraffin wax embedded tissues. J Clin Pathol 1991; 44: 924-927.
18. Longy M, Duboue B, Soubeyran P, Moynet D. Method for the purification of tissue DNA suitable for PCR after fixation with Bouin’s fluid. Uses and limitations in microsatellite typing. Diagn Mol Pathol 1997; 6: 167-173.
19. Sato V, Sugie R, Tsuchiya B, Kameya T, Natori M, et al. Comparison of the DNA extraction methods for polymerase chain reaction amplification from formalin-fixed and paraffin-em bedded tissues. Diagn Mol Pathol 2001; 10: 265-271.
20. Tbakhi A, Totos G, Pettay Jd, Myles J, Tubbs Rr. The effect of fixation on detection of B-cell c1onality by polymerase chain reaction. Mod Pathol 1999; 12: 272-278.
21. Goelz Se, Hamilton Sr, Vogelstein B. Purification of DNA from formaldehyde fixed and paraffin embedded human tissue. Biochem Biophys Res Commun 1985; 130: 118-126.
22. Tvrdik, D., Svatošova, J., Dundr, P., Povyšil, C. Molecular Diagnosis of Synovial Sarcoma: Detection of SYTSSX1/ 2 Fusion Transcripts by RT-PCR in Paraffin-Embedded Tissue. Med. Sci. Monit. 2005; 11:1-7.