Sensitivity of an immunoglobulin heavy chain gene polymerase chain reaction primer system for routine diagnosis of lymphomas

  • R Heyny-von Haußen1,

    Affiliated with

    • C Braun1 and

      Affiliated with

      • G Mall1

        Affiliated with

        Diagnostic Pathology20072(Suppl 1):S7

        DOI: 10.1186/1746-1596-2-S1-S7

        Published: 14 March 2007

        Aims

        Polymerase chain reaction (PCR) and length fragment analyses of the immunoglobulin heavy chain (IgH) gene are useful tools for clonality assessment in malignant B-cell-lymphomas and reactive lymphoid infiltrates. The present investigation analyzes the sensitivity of an IgH gene consensus primer multiplex PCR system for the detection of clonality for routine diagnosis in 109 lymphomas during a period of 3 years (2003–2005) at the Institut of Pathology Klinikum Darmstadt.

        Methods

        We used FR2A/JH/VLJH and FR3A/JH/VLJH primer sets for detecting clonal B cell populations. Primer sequences used for PCR: Variable region FR2A consensus primer: FR2A: TGG(AG)TCCG(AC)CAG(GC) C(CT)(CT)C(AGCT)GG. Joining region (JH) consensus primer: LJH: TGAGGA GACGGTGACC, VLJH: GTGCAGGT(AGCT) CCTTGGCCCCAG-FAM. 1. PCR: FR2A/LJH and FR3A/LJH. 2. PCR: FR2A/VLJH-FAM and FR3A/VLJH-FAM. Fluorescence fragment analyses of IgH gene rearrangement were performed with FAM-labeled PCR products by high-resolution capillary electrophoresis using the ABI-PRISM 310 Genetic Analyzer (Applied Biosystems, Weiterstadt, Germany) with a POP 6-filled capillary (Applied Biosystems) and analyzed by using the GeneScan software (Applied Biosystems). A fragment was considered to be clonal in the case of a peak-height ratio (PHR) >2. The PHR was calculated as the quotient of the highest peak divided by the mean height of the two peaks surrounding the largest peak.

        Results

        See Table 1.
        Table 1

         

        Lymphoma diagnosis

        Detection of clonally rearranged IgH gene

        B lymphoblastic lymphoma

        100% (1/1)

        Mantle cell lymphoma

        100% (3/3)

        CLL

        100% (24/24)

        Lymphoplasmacytic lymphoma

        100% (6/6)

        Marginal zone B-cell lymphoma

        100% (48/48)

        Plasma cell myeloma

        100% (1/1)

        Hairy cell leukaemia

        50% (1/2)

        Follicular lymphoma

        55% (6/11)

        Diffuse large B-cell lymphoma

        82% (9/11)

        Hodgkin lymphoma

        50% (1/2)

        Conclusion

        The present study showed a variable sensitivity of PCR of IgH gene region with FR2A/JH/VLJH and FR3A/JH/VLJH consensus primer sets for different lymphomas. Sensitivity of our consensus primer set mainly depends on the existence of IgH gene rearrangements or the status of the IgH gene in the special lymphoma types, especially the occurrence of somatic hypermutation in variable-region genes.

        Authors’ Affiliations

        (1)
        Pathologisches Institut Klinikum Darmstadt

        Copyright

        © Heyny-von Haußen et al 2007

        This article is published under license to BioMed Central Ltd.