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The utility of the BIOMED-2 primers in the detection of 2 clonal, B-lymphoproliferative disorders simultaneously involving the same site.

Molecular studies are widely used in the diagnosis of lymphomas. (1-3) Tests by polymerase chain reaction (PCR) using DNA extracted from the specimen as a template and primers for immunoglobulin genes are probably among the more commonly ordered molecular tests. These studies are used to identify clonal populations in specimens with equivocal morphology or immunopheno-type. (1,3,4) In many cases, the diagnosis relies heavily on the sensitivity and specificity of the molecular tests, and considerable effort has been made to develop a set of primers that allows reliable identification of clonal rearrangements. (2,3) The BIOMED-2 Concerted Action BMH4CT98-3936 has developed a set of guidelines and primers that are currently used widely. Those primers have been tested at multiple institutions and have demonstrated their efficacy in the detection of clonal populations. (1-4)

In the past few decades, significant advances have been made in the diagnosis, classification, and treatment of lymphoproliferative disorders. (5) Given the therapeutic options available, an accurate diagnosis of lymphoid lesions is critical. Monoclonal B-cell lymphocytosis (MBL), an indolent lymphoproliferative disorder, is thought to be present in up to 10% of the elderly (6-8) and, as expected, can be responsible for a positive PCR test result when conducted on a specimen suspicious for lymphoma, even if the main lesion is not involved by lymphoma. The ability of the test to diagnose 2 clonal proliferations in the same sample is essential in this type of situation because the positive clonal result may be a consequence of MBL, but it could be interpreted as evidence of a different, possibly more aggressive, lymphoproliferative disorder. Differentiating between 1 or 2 clonal populations in the same sample may also be crucial when a clonal lymphoid population is identified simultaneously with a population that has plasmacytoid features. If both cell populations are part of the same clone they could represent marginal zone lymphoma or lymphoplasmacytic lymphoma, (9,10) whereas if they are separate clones, they may indicate a low-grade B-cell lymphoma and a concurrent plasma-cell neoplasm, such as monoclonal gammopathy of undetermined significance. (11) Obviously, the management of the 2 types of cases is very different.

Recent studies have clearly demonstrated the ability of the BIOMED-2 primers to generate multiple amplicons in the same reaction. (12,13) To our knowledge, however, there are few reliable studies investigating the ability of the BIOMED-2 primers to identify 2 cell populations with clonally rearranged immunoglobulin genes in the same specimen. To investigate the utility of these primers in a clinical setting, we selected 10 formalin-fixed, paraffin-embedded specimens in which 2 clonal populations had been diagnosed by flow cytometry, morphology, or immunohistochemistry. To avoid the possibility that both populations were part of the same clone, cases of diffuse large B-cell lymphoma (DLBCL) arising in a background of follicular lymphoma (FL) or marginal zone lymphoma (MZL) were precluded if no other B-cell lymphoproliferative disorders were present. There were 3 cases (30%) of FL associated with chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL) or MBL, 3 cases (30%) of lymphoplasmacytic lymphoma/ MZL and CLL/MBL, 3 cases (30%) of CD5- DLBCL associated with CLL/MBL, and 1 case (10%) of FL and DLBCL associated with MBL.


Case Selection

After institutional review board approval, cases were selected from the archives of the Pathology and Laboratory Medicine Institute (Cleveland Clinic, Cleveland, Ohio). The diagnosis was confirmed by histomorphologic examination in all cases. In 9 of the 10 cases (90%), flow cytometric data were also available. Immunohistochemical stains were available in all 10 cases (100%), including the case not analyzed by flow cytometry. In addition, 94 consecutive molecular tests that were interpreted as positive for a clonal amplicon were identified, and the data were reviewed.

Immunohistochemical Stains

Immunohistochemical stains were performed on 4-[micro]m-thick sections of formalin-fixed, paraffin-embedded tissue, using an automatic immunostainer (Benchmark XT or Ultra, Ventana Medical Systems, Tucson, Arizona). The antibodies anti-CD3 (Cell Marque, Rocklin, California; catalog No. CMC365), CD5 and CD10 (Novocastra, Newcastle Upon Tyne, UK; catalog No. NCL-CD10-279), CD20 (Dako, Carpinteria, California; catalog No. M0755), CD21 (Dako; catalog No. M0784), CD79a (Dako; catalog No. M7050), cyclin D1, BCL2 (Cell Marque, catalog No. 760-4240), BCL6 (Dako, catalog No. M7211), MUM1 (Dako, catalog No. 7259), PAX5 (BD Biosciences, San Jose, California; catalog No. 610863), free [kappa] light chain (Dako, catalog No. A0100), free [lambda] light chain (Dako, catalog No. A0101) were used. Detection of protein-antibody complexes was achieved with an inobenzidine tetrahydrochloride detection kit (iVIEW DAB Detections, Ventana; catalog No. 760-091).

DNA Extraction

There was no microdissection or selection of areas to be analyzed and no cell sorting. DNA from the formalin-fixed, paraffin-embedded tissue was extracted using a High Pure PCR Template Preparation Kit (Roche Applied Science, Indianapolis, Indiana; catalog No. 11796828001). The tissue for DNA extraction was obtained from consecutive sections of the paraffin blocks. The first and last sections were stained with hematoxylin-eosin, to ensure that the entire lesional tissue was represented in the sections submitted for DNA extraction and PCR.

Polymerase Chain Reaction

The PCR tests were conducted in duplicate, using DNA extracted from the paraffin blocks as templates. The polymerase used was AmpliTaq Gold with the appropriate buffer (Applied Biosystems, Carlsbad, California; catalog No. 4311814). The quality of the purified DNA was verified using a Specimen Control Size Ladder6FAM (Invivoscribe, San Diego, California; catalog No. 2-096-0021). In addition to the patient samples, each run included reactions using Human Polyclonal Genomic DNA from Tonsil (Invivoscribe; catalog No. 4-092-0010) and the Sensitivity Panel IVS-0019 Control Clonal DNA (Invivoscribe; catalog No. 4-086-0190) as templates. The following fluorescently labeled primers were used: IGH FR1-6FAM (Invivoscribe; catalog No. A-101-0011), IGH FR2-6FAM (Invivoscribe; catalog No. A-101-0101), IGH FR3- HEX (Invivoscribe; catalog No. A-101-0031), IGK V-J-6FAM (Invivoscribe; catalog No. A-102-0011), and IGK V-Kde-6FAM (Invivoscribe; catalog No. A-102-0021). After the specimens were unloaded from the thermocycler, the results of the PCRs were analyzed with a 3730xl DNA Analyzer (Applied Biosystems), using the protocols recommended by the manufacturer. The data acquired were analyzed with GeneMapper software (Applied Biosystems).

Interpretation of the PCR Data

Tracings from all the reaction tubes were reviewed. Peaks that were considered clonal amplicons were reproducible, dominant (height at least 2-fold that of the peaks in the background), and within the size acceptable for the primers used. Low-amplitude peaks, of a height equal or smaller than that of the polyclonal background were not considered. When the reaction generated 2 dominant peaks of very close size, within 5 nucleotides, and of identical height, they were interpreted to indicate only one clonal population. These criteria were applied consistently to all the specimens examined.


In most cases, histologic examination and medical history aroused suspicion for a collision neoplasm. In addition to morphology, immunohistochemistry contributed to the identification of the lymphoproliferative disorders in the specimens (Figure 1, A through F). Flow cytometric data were available on 9 of the 10 cases (90%) (Table), which demonstrated 2 abnormal lymphoid populations in 7 out of those 9 cases (78%) (Table; Figure 2). In the 2 cases (22%) with only one abnormal population by flow cytometry, that population was positive for CD5, whereas the morphologic and immunohistochemical features showed the presence of CD[10.sup.+] FL or DLBCL. In 5 out of the 7 specimens (71%) with 2 clonal populations by flow cytometry, one abnormal clone was positive for [kappa] surface immunoglobulin light chains, whereas the other was positive for [lambda] light chains (Figure 2). In a case with a diagnosis that included MBL, one of the populations was negative for surface immunoglobulin light chains and positive for CD5, whereas the other population was monotypic for k light chains. In one case diagnosed as CLL and FL, both clonal populations were positive for [lambda] surface immunoglobulin light chains, but one was positive for CD5, whereas the other was positive for CD10 (Table). In the only case in which flow cytometric data were not available, immunohistochemistry demonstrated that, in addition to the CD[10.sup.+] B-cell population diagnosed as FL, there was a CD[5.sup.+] B-cell population (Figure 1, A through F). These findings and the clinical history supported the diagnosis of 2 clonal populations involving the same site. Extensive immunohistochemical characterization of the cells was available in all 10 cases (100%).

In 9 out of the 10 cases (90%), there was amplification in all 5 tubes (Table). In one case (10%), there was no amplification in the tubes with primers for IGH FR1 and IGH FR2. However, in all 10 specimens (100%), at least one of the primer combinations showed an amplification product, indicating the presence of a population with clonally rearranged immunoglobulin genes. No case (0%) had 2 clonal peaks with all 5 or with 4 of the primer combinations tested (IGH FR1, IGH FR2, IGH FR3, IGK VJ, and IGK VKde) (Figure 3). Only one case (10%) showed 2 peaks with 3 primer sets. In 4 cases (40%), 2 clonal amplicons were detected in 2 reactions. A similar result was detected with only one primer set in 3 cases (30%). Overall, 8 out of the 10 specimens (80%) displayed 2 clonal peaks with at least one primer combination. One of the cases (10%) that showed only one clonal population by PCR was the case that showed no amplification with 2 primer combinations, and the other case (10%) had good amplification with all primer combinations.


As expected, not all primer combinations were equally effective in the detection of 2 clonal populations in the same specimen. In 4 out of the 10 cases (40%) tested (or 4 of the 9 cases [44%] with amplification) the primers for IGH FR2 showed 2 clonal peaks (Table). Primers for IGK VJ and IGK VKde showed 2 amplification peaks in 3 cases each (30% each). Interestingly, only one specimen (10%) demonstrated 2 peaks with both IGK VJ and IGK VKde primer sets. In one case (10%), positivity with 1 of those 2 primer sets was the only molecular evidence of biclonality; 2 clonal populations were identified with IGH FR1 and IGH FR3 primer sets in 4 cases (40%; 2 [20%] with IGH FR1 and 2 [20%] with IGH FR3), and 2 of these cases (the ones with biclonal results for IGH FR1) also demonstrated biclonal populations with the IGH FR2 primer set. One of the biclonal cases (10%) by IGH FR3 was biclonal with IGK VJ. One of the cases (10%) demonstrated biclonality with the IGH FR3 primer set only.

To summarize the results, in 8 out of the 10 cases (80%), the 2 clonal processes were identified by molecular methods. The most effective primer combination was IGH FR2, which was positive in 4 of the 10 cases (40%) tested. The least-useful primers were those for IGH FR1, which showed positive results in only 2 out of the 10 specimens (20%), but in neither of those 2 specimens were they the only positive results.


Review of the data from 94 consecutive PCR tests that had been interpreted to be positive showed that multiple clonal amplicons were detected in 10 cases (10.6%). In all these 10 cases (100%), 2 dominant amplicons were detected in only one reaction. In 2 cases (20%), the difference in size between the amplicons was less than 5 nucleotides, and the 2 peaks had identical heights, an indication that they were the result of the same neoplastic B-cell clone. Whether the 2 amplicons detected in 8 cases (80%) were consequences of biclonal processes or of biallelic rearrangements is unclear. Examination of the routine sections from these specimens did not raise suspicion for 2 lymphoproliferative disorders. There were no flow cytometric data on these cases.


Rapidly accumulating data on the pathobiology of the hematolymphoid neoplasms have resulted in the recognition of new diagnostic categories. (5) As targeted therapies are continuously being generated, establishing the precise diagnosis in each case is important for more than just academic or research purposes. The increasing sensitivity of diagnostic methods, which can result in detection of neoplasms at very early stages when they do not display all the diagnostic morphologic or immunophenotypic features, further complicates matters. Moreover, with the aging population, the number of patients who may have unrecognized MBL or monoclonal gammopathy of undetermined significance is continuously increasing. (5,6,8,11) Just by chance alone, some of these patients may develop other types of lymphoma or they may acquire atypical inflammatory infiltrates. Using amplification methods in the investigation of atypical lymphoid proliferations in patients who may have other clonal abnormalities (MBL or monoclonal gammopathy of undetermined significance) may lead to diagnostic errors, because the positive clonal molecular result, secondary to MBL or monoclonal gammopathy of undetermined significance, may be attributed to a possibly benign lesion.


The observation that, in lymphoid malignancies, the abnormal cells have the immunoglobulin genes or the T-cell receptor genes clonally rearranged has led to the development of molecular tests that investigate the rearrangement status of these genes. (2) Tests using Southern blot techniques were among the first molecular tests in clinical use. (14) However, these labor-intensive, time-consuming tests often require fresh-frozen tissue, which severely limits their clinical use. Some of the reagents are labeled with radioactive isotopes, which discourages many laboratories to develop Southern blot assays. To circumvent these limitations, PCR was developed. (15) In time, collaboration between institutions led to the BIOMED-2 consensus. (2) The methods and primers developed in that process are now considered the gold standard. (1,3) The sensitivity of those primers has been tested by mixing clonal cell populations with polyclonal, normal lymphocytes. The ability of the BIOMED-2 primers to detect a second clonal population in the presence of a dominant population has not been studied extensively. Moreover, experiments on artificial dilutions of clonal populations in the laboratory may not reflect the performance of the tests on actual patient samples, when the ratio between different cell populations (between clones and/or normal, polyclonal cells) cannot be controlled.

In clinical practice, certain diagnostic dilemmas cannot be resolved reliably in the absence of data regarding the efficiency of the BIOMED-2 primers. For example, flow cytometry or immunohistochemistry may identify a monotypic, CD5- B-cell population along with a small clonal plasma-cell population in the same bone marrow specimen. If the B cells and the plasma cells are part of the same clone, the diagnosis of lymphoplasmacytic lymphoma or MZL is warranted. If the plasma cells constitute a separate clone in addition to the lymphoproliferative disorder, a plasma cell neoplasm should also be diagnosed. The management and prognosis of those 2 diagnostic possibilities are radically different.

We have investigated the ability of the BIOMED-2 primers to detect 2 clonal, B-cell populations in the same sample. We identified samples in which the 2 different populations were diagnosed by morphology, flow cytometry, and immunohistochemistry. To avoid the possibility that the 2 populations were part of the same clone, we did not select cases of DLBCL arising from FL or MZL, with the exception of one case, in which, in addition to FL and DLBCL, there also was a CD5+, clonal B-cell population. In all the cases, one of the components was MBL or CLL, a lymphoproliferative disorder characterized by positivity for CD5. The other clone was positive for CD10 (in cases with FL or DLBCL) or negative for CD5 and had strong surface expression of CD20, CD79b, and FMC7 (in MZL). The ratio between the 2 clonal populations or between the abnormal populations and the normal cells in the background was difficult to assess. Moreover, whether the proportion of the 2 clones present in the same sample analyzed by PCR was similar to that analyzed by flow cytometry or morphology was unclear. Although this inability to precisely quantify the clonal populations may be perceived as a limitation, we actually interpreted it as a strength of this study because, in routine clinical practice, it is difficult to obtain that information on most specimens; hence, the current study closely reproduces the use of molecular tests in practice.

As expected, different primer combinations showed varying efficiencies in the detection of clonal populations. In 2 reactions, there was no amplification product. Only 4 of the 48 reactions (8%) with adequate amplification products showed no evidence of a clonal rearrangement and, in 3 of those 4 reactions (75%), the primers used were for IGK VKde. The first conclusion was that, in our hands, the BIOMED-2 protocol detected one clonal population with an efficiency similar to that previously reported in the literature. (1,3)

The identification of 2 clonal populations in 8 of the 10 cases (80%) tested shows that, in many cases, the existence of 2 clones in the same specimen can be detected. In 1 of the 2 cases (50%) in which we could identify only one clonal amplicon, 2 of the 5 reactions (40%) failed to generate any amplification products, but 2 of the remaining tubes (40%) showed a clonal product. In most cases, only one or 2 reactions generated 2 dominant amplicons. The IGH FR2 primer combination was the most effective in the detection of 2 clonal populations. Interestingly, the combinations for IGK VJ and IGK VKde were more effective than were the primers for IGH FR1 or IGH FR3. In all the cases with 2 clonal amplification products in the same tube, the difference in size between the amplicons was greater than a few nucleotides, making the possibility that the separate amplicons were the result of somatic hypermutation unlikely. The presence of 2 clonal amplicons was useful in the diagnosis of the second lymphoproliferative disorder, but it provided no information beyond that point because further refining of the diagnosis was the result of morphologic, immunohistochemical, and flow cytometric analysis.

As suggested in the peer-review process, there was a distinct possibility that the presence of multiple clonal amplicons was the consequence of biallelic rearrangements and that the multiclonal result was actually generated by only one of the clonal populations in the sample. The effect of biallelic arrangements on the results of PCR tests is often ignored in the literature and in practice, but recent works demonstrate that sensitive, well-designed tests should be able to identify biallelic rearrangements in a large proportion of the cases. (12,13) To determine the sensitivity of our methods and criteria in the identification of biallelic rearrangements, we analyzed the data from 94 consecutive tests that had been diagnosed as positive and in which there was no stated suspicion for a biclonal process. Approximately 10% of these cases (n = 9) showed 2 amplicons, but in 2% of the cases (n = 2), the pattern of the amplification peaks clearly indicated that they were part of the same clone. Moreover, 2 amplicons were detected in only 1 of the 5 reactions (20%) conducted in each case. These findings were in sharp contrast with the data from the specimens in which the presence of 2 lymphoproliferative disorders had been proven by morphologic and immunophenotypic examination. A useful finding for differentiating biclonal and biallelic processes is the number of amplicons detected with the primers sets for IGK VJ and IGK VKde. If there are more than 2 amplicons (3 or 4) being generated by each of these primer combinations, it is likely that the specimen contains a biclonal process.

There are several important implications of this study. First, reporting the results of the BIOMED-2 in aggregate as positive or negative is inadequate. Reporting of the results for each primer combination would allow a more-detailed description of the findings. The number of dominant amplicons should be mentioned in the result for each primer combination, especially for differentiating biallelic and biclonal processes, which may depend on that information. Second, the detection of 2 dominant amplicons may lead to either the de novo diagnosis of 2 clonal processes or to the identification of a second clonal process (in patients with a history of a lymphoproliferative or plasma cell disorders). Third, the interpretation of the results of the BIOMED-2 tests has to consider the medical history and morphologic data, especially when a biclonal or biallelic process is detected by molecular techniques. Fourth, special attention should be devoted to the examination of data generated with primers for IGH FR2, IGK VJ, or IGK VKde.

In conclusion, the BIOMED-2 primers are a useful tool in the diagnosis of multiple, clonal, B-cell populations in the same sample. Similar studies conducted at several institutions could lead to the development of criteria that reliably differentiate between biclonal and biallelic processes.

Caption: Figure 1. Immunohistochemistry in a case of follicular lymphoma and chronic lymphocytic leukemia/small lymphocytic lymphoma. A, The B cells with extrafollicular distribution show a dimmer staining pattern than do the follicular B cells. B, T cells, mainly extrafollicular. C, The follicle is positive for CD10. D, The follicular cells are also positive for BCL6. E, Most extrafollicular cells are positive for CD5, with a 2-intensity pattern: The T cells are strongly positive, whereas most cells, corresponding to the B cells (compare with A and B), are dimly positive. F, All the cells are positive for BCL2, but the follicular cells have a strong staining pattern, probably a consequence of t(14;18) (CD20, original magnification X10 [A]; CD3, original magnification X10 [B]; CD10, original magnification X10 [C]; BCL6, original magnification X10 [D]; CD5, original magnification X10 [E]; and BCL2, original magnification X10) (F).

Caption: Figure 2. Flow cytometry. CD45 and light side-scatter gating show CD5+ T cells (red) and CD19+ and CD20+ B cells (blue and green). There are 2 populations of B cells: one is positive for CD5, monotypic lambda light chains (green), and the other is negative for CD5, monotypic kappa light chains (blue).

Caption: Figure 3. Example of a biclonal result: 2 independent peaks of different amplitudes and sizes--148 and 206 nucleotides, in duplicate, on a polyclonal background (IGK VJ primer set).


(1.) Evans PA, Pott C, Groenen PJ, et al. Significantly improved PCR-based clonality testing in B-cell malignancies by use of multiple immunoglobulin gene targets. Report of the BIOMED-2 Concerted Action BHM4-CT98-3936. Leukemia. 2007;21(2):207-214.

(2.) van Dongen JJ, Langerak AW, Bruggemann 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.

(3.) van Krieken JH, Langerak AW, Macintyre EA, et al. Improved reliability of lymphoma diagnostics via PCR-based clonality testing: report of the BIOMED-2 Concerted Action BHM4-CT98-3936. Leukemia. 2007;21(2):201-206.

(4.) Langerak AW, Molina TJ, Lavender FL, et al. Polymerase chain reaction-based clonality testing in tissue samples with reactive lymphoproliferations: usefulness and pitfalls. A report of the BIOMED-2 Concerted Action BMH4CT98-3936. Leukemia. 2007;21(2):222-229.

(5.) Swerdlow SH, Campo E, Harris NL, et al, eds WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. 4th ed. Lyon, France: IARC Press; 2008. World Health Organization Classification of Tumours; vol 2.

(6.) Ghia P, Prato G, Scielzo C, et al. Monoclonal CD5+ and CD5 B-lymphocyte expansions are frequent in the peripheral blood of the elderly. Blood. 2004;103(6):2337-2342.

(7.) Han T, Ozer H, Gavigan M, et al. Benign monoclonal B cell Lymphocytosis--a benign variant of CLL: clinical, immunologic, phenotypic, and cytogenetic studies in 20 patients. Blood. 1984;64(1):244-252.

(8.) Rawstron AC, Bennett FL, O'Connor SJ, et al. Monoclonal B-cell lymphocytosis and chronic lymphocytic leukemia. N Engl J Med. 2008;359(6): 575-583.

(9.) Coupland SE, Hellmich M, Auw-Haedrich C, Lee WR, Anagnostopoulos I, Stein H. Plasmacellular differentiation in extranodal marginal zone B cell lymphomas of the ocular adnexa: an analysis of the neoplastic plasma cell phenotype and its prognostic significance in 136 cases. Br J Ophthalmol. 2005; 89(3):352-359.

(10.) Lin P, Medeiros LJ. Lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia: an evolving concept. Adv Anat Pathol. 2005;12(5):246-255.

(11.) Blade J, Rosinol L, Cibeira MT, de Larrea CF. Pathogenesis and progression of monoclonal gammopathy of undetermined significance. Leukemia. 2008; 22(9):1651-1657.

(12.) Langerak AW, van Dongen JJM. Multiple clonal Ig/TCR products: implications for interpretation of clonality findings. J Hematopathol. 2012;5(1 2):35-43.

(13.) Langerak AW, Groenen PJ, Bruggemann M, et al. EuroClonality/BIOMED-2 guidelines for interpretation and reporting of Ig/TCR clonality testing in suspected lymphoproliferations. Leukemia. 2012;26(10):2159-2171.

(14.) Cossman J, Zehnbauer B, Garrett CT, et al. Gene rearrangements in the diagnosis of lymphoma/leukemia: guidelines for use based on a multiinstitutional study. Am J Clin Pathol. 1991;95(3):347-354.

(15.) Horsman DE, Gascoyne RD, Coupland RW, Coldman AJ, Adomat SA. Comparison of cytogenetic analysis, southern analysis, and polymerase chain reaction for the detection of t(14;18) in follicular lymphoma. Am J Clin Pathol. 1995;103(4):472-478.

Brenda Ly, MD; Claudiu V. Cotta, MD, PhD

Accepted for publication February 7, 2013.

From the RJ Tomsich Pathology and Laboratory Medicine Institute, Cleveland Clinic, Cleveland, Ohio.

The authors have no relevant financial interest in the products or companies described in this article.

Reprints: Claudiu V. Cotta, MD, PhD, RJ Tomsich Pathology and Laboratory Medicine Institute, Cleveland Clinic, 9500 Euclid Ave/L11, Cleveland, OH 44195 (e-mail:
Immunophenotypic and Molecular Data

Diagnosis                  Light          Methods        IGH FR1,
                           Chains                           No.

FL + CLL                     LL       Flow + IHC             0
MZL + CLL                    KL       Flow + IHC             2
FL + MBL                              IHC + history          1
LPL + MBL                    K0       Flow + IHC             1
MZL + CLL                    KL       Flow + IHC             2
DLBCL + CLL                  KL       Flow + IHC             1
FLis + MBL                    0       Flow + IHC             1
DLBCL + CLL                  KL       Flow + IHC             1
DLBCL + FL + MBL              L       Flow                   1
DLBCL + MBL                  KL       Flow                  N/A
Biclonal cases, No.                                          2

Diagnosis                  IGH FR2,      IGH FR3,      IGK VJ,
                              No.           No.          No.

FL + CLL                       1             2            2
MZL + CLL                      2             1            1
FL + MBL                       1             2            1
LPL + MBL                      1             1            1
MZL + CLL                      2             1            1
DLBCL + CLL                    1             1            2
FLis + MBL                     2             1            2
DLBCL + CLL                    2             1            1
DLBCL + FL + MBL               1             1            1
DLBCL + MBL                   N/A            1            1
Biclonal cases, No.            4             2            3

Diagnosis                  IGK Vkde,

FL + CLL                  1
MZL + CLL                 1
FL + MBL                  0
LPL + MBL                 2
MZL + CLL                 2
DLBCL + CLL               2
FLis + MBL                0
DLBCL + CLL               1
DLBCL + FL + MBL          1
DLBCL + MBL               0
Biclonal cases, No.       3

Abbreviations: 0, amplified, but no clonal peak;1, 1 clonal peak;
2, 2 clonal peaks; CLL, chronic lymphocytic leukemia/small
lymphocytic lymphoma; DLBCL, diffuse large B-cell lymphoma; FL,
follicular lymphoma; FLis, follicular lymphoma in situ; Flow, flow
cytometry; IHC, immunochemistry; K, [kappa] immunoglobulin light
chains; L, [lambda] immunoglobulin light chains; LPL,
lymphoplasmacytic lymphoma; MBL, monoclonal B lymphocytosis; MZL,
marginal zone lymphoma; N/A, not amplified.
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Author:Ly, Brenda; Cotta, Claudiu V.
Publication:Archives of Pathology & Laboratory Medicine
Article Type:Report
Geographic Code:1USA
Date:Nov 1, 2013
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