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PCR Analysis of IgH and TCR-γ Gene Rearrangements as a Confirmatory Diagnostic Tool for Lymphoproliferative Disorders

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Abstract

This study investigates PCR analysis of immunoglobulin heavy chain (IgH) and T cell receptor (TCR) gene rearrangements on paraffin-embedded tissue sections and bone marrow aspirates of patients suspected to have lymphoproliferative disorders but with inconclusive diagnosis in histopathological examination. 130 samples of patients with inconclusive immunohistochemistry results were evaluated for clonal rearrangement of IgH and TCR genes. Based on histopathology examination, the patients were divided into three groups: the first group without any definite diagnosis of lymphoproliferative disorders (60 cases, 46.2 %), the second group suspected to have a lymphoproliferative disorder but in favor of benign disorders (19 cases, 14.6 %) and the third group suspect to lymphoproliferative disorders but relatively in favor of malignant disorders (51 cases, 39.2 %). After DNA extraction and quality control, semi-nested PCR was performed using consensus primers for amplification of TCR-γ and CDR-3 regions of IgH genes. PCR products were analyzed after heteroduplex analysis using polyacrylamide gel electrophoresis, and were subject to silver staining. Totally, in over half of the cases (55.4 %), a monoclonal pattern was found in IgH or TCR-γ genes rearrangements. Monoclonal IgH gene rearrangement was detected in 48.1 % of patients, whereas monoclonal TCR-γ gene rearrangement was found in 33.6 % of them, which was not statistically significant (P = 0.008). Only in 32 patients (24.6 %) were the results of TCR-γ and IgH gene rearrangements consistent with respect to the presence (2.3 %) or absence (22.3 %) of monoclonality. Finally, PCR analysis of TCR-γ and IgH gene rearrangements led to definite diagnosis in 105 patients (80.8 %), and only 25 cases (19.2 %) remained inconclusive. Our results emphasize the usefulness of gene rearrangement study in cases without a definite diagnosis in immunohistochemistry studies. Multiple PCR analysis results when combined with patient’s clinical course and immunohistochemistry can lead to early diagnosis and subsequent therapy.

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Acknowledgments

We wish to thank all our colleagues in Payvand Clinical and Specialty Laboratory.

Conflict of Interest

None of the authors has conflict of interest.

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Authors and Affiliations

  1. Tehran Medical Branch, Islamic Azad University, Zargandeh St, Shariati Ave, Tehran, Iran

    Behzad Poopak

  2. Students’ Scientific Research Center (SSRC), Tehran University of Medical Sciences (TUMS), Tehran, Iran

    Ali Kord Valeshabad

  3. National Foundation of Elites, Tehran, Iran

    Ali Kord Valeshabad

  4. Cancer Institute of Imam Khomeini Hospital, Tehran Medical Sciences University, Tehran, Iran

    Fazel Elahi

  5. Hemato-Oncology Ward, Taleghani Hospital, Shahid Behshti Medical Sciences University, Tehran, Iran

    Hamid Rezvani

  6. Payvand Clinical & Specialty Laboratory, No. 174 Zafar St., Shariati Ave, Tehran, Iran

    Gelareh Khosravipour, Mohammad Ali Jahangirpour, Shirin Bolouri & Tolou Golkar

  7. Health Research Institute, Research Center of Thalassemia and Hemoglobinopathy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

    Fatemeh Salari, Mohammad Shahjahani & Najmaldin Saki

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  1. Behzad Poopak
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  11. Najmaldin Saki
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Correspondence to Behzad Poopak.

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Poopak, B., Valeshabad, A.K., Elahi, F. et al. PCR Analysis of IgH and TCR-γ Gene Rearrangements as a Confirmatory Diagnostic Tool for Lymphoproliferative Disorders. Indian J Hematol Blood Transfus 31, 38–45 (2015). https://doi.org/10.1007/s12288-014-0387-z

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  • DOI: https://doi.org/10.1007/s12288-014-0387-z

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